Probe combination, kit and method for detecting gene variation of dmd, dmd modified gene and differential diagnosis

Abstract

The present invention relates to a method for detecting DMD a genetic, DMD modification gene and a probe combination, kit and method for differentiating gene variation. The present invention particularly relates to a probe combination comprising the following 6 probe groups (1) and (6), or (1), (2), (3) and (6), or (1), (2), (3), (4), (5) and (6): (1) a probe group targeting a DMD gene region; (2) a probe group targeting a DMD gene regulatory region; (3) a probe group targeting a DMD DMD modification gene region; (4) a probe group targeting a differentiating gene region; (5) a probe group targeting a mitochondrial gene region; (6) a probe group targeting a gender identification gene region.

Description

Technical Field

[0001] This invention relates to the field of gene detection, specifically to the detection of... DMD Gene, DMD Probe combinations, kits, and methods for modifying genes and identifying gene variations. Background Technology

[0002] Duchenne Muscular Dystrophy (DMD) is an X-linked recessive genetic disorder caused by mutations in the dystrophin gene. The disease-causing gene is... DMD The gene, located in the Xp21 region of the X chromosome, is the largest known gene in the human genome, containing 79 exons. Its complex structure and large sequence span are noteworthy. Dystrophin plays a crucial role in maintaining the structural integrity of muscle cell membranes. Its absence leads to decreased muscle cell membrane stability, resulting in repeated muscle fiber damage and repair failure, ultimately causing progressive muscle weakness and loss of motor function. It can also be accompanied by involvement of the myocardium and respiratory system, severely impacting patients' quality of life and lifespan.

[0003] Duchenne muscular dystrophy (DMD) has a high incidence rate, ranging from approximately 1 in 3,500 to 1 in 5,000 male newborns. The disease exhibits significant genetic heterogeneity, with about two-thirds of cases inherited maternally and the remaining one-third caused by de novo mutations. This indicates a high rate of spontaneous mutation in the gene, making genotype prediction crucial for understanding the pathogenesis of DMD.

[0004] Regarding molecular pathogenic mechanisms, DMD Gene mutations are complex and diverse, mainly including large deletions, duplications, and small mutations (including point mutations, small insertions or deletions, and splice site mutations). Exon deletions are the most common type of mutation, accounting for approximately 60%–70%; exon duplications account for approximately 5%–15%; and the remaining 20%–30% are small mutations. Among small mutations, nonsense mutations, frameshift mutations, and splice site mutations constitute the majority. Furthermore, DMD The gene exhibits distinct mutation hotspots, primarily concentrated in exons 2–19 and 45–55. These mutation types and distribution characteristics increase the complexity of molecular detection and place higher demands on the sensitivity and coverage of detection technologies.

[0005] In addition to the common mutation types mentioned above, DMD Pathogenic variants also exist in genes located in deep intron regions. These variants typically affect normal protein expression by activating aberrant splicing sites to form pseudoexons. Existing research indicates that these variants... DMDWhile the proportion of mutations in these cells is approximately 1%–3%, the detection rate can reach 5%–10% or more in patients who cannot be definitively diagnosed by routine testing. Because traditional detection methods struggle to cover deep intron regions, these mutations are easily missed, making them one of the major technical blind spots in the current molecular diagnosis of Duchenne muscular dystrophy.

[0006] against DMD Molecular detection techniques for exon-level deletions and duplications primarily include multiplex ligation-dependent probe amplification (MLPA), next-generation sequencing (NGS) based on high-throughput sequencing, and third-generation sequencing methods. Among these, MLPA is mainly used to detect exon-level deletions and duplications and is one of the most widely used clinical methods. This method can detect... DMD Relative quantitative analysis of all exons of a gene offers advantages such as ease of operation, high detection efficiency, and relatively low cost. However, this method is only suitable for detecting copy number variations and cannot identify point mutations or small insertions / deletions. Furthermore, it is susceptible to variations in the probe binding region sequence in cases of single-exon abnormalities, potentially leading to false positives or false negatives. In addition, MLPA has significant technical limitations in detecting complex structural variations such as inversions and translocations. NGS, on the other hand, can achieve... DMD High-throughput sequencing of gene coding regions can simultaneously detect point mutations, small insertions / deletions, and partial copy number variations, thereby improving mutation detection rates. However, existing NGS methods still have limitations in detecting large-scale copy number variations, especially in the identification of repetitive sequence regions or complex structural variations. With the development of third-generation sequencing technology, third-generation capture-based techniques have also been applied. DMD The testing is underway, but mainly focuses on... DMD Gene mutation analysis focuses primarily on deletions, duplications, and small mutations, neglecting the prediction and analysis of deep intron variations. All of these techniques suffer from limitations in explaining patient clinical phenotypic differences, hindering prognostic assessment, and making them susceptible to confusion with other hereditary myopathy disorders.

[0007] Introduction DMD Detection of modified genes can reveal genetic factors influencing disease progression at the molecular level, enabling the assessment of disease severity and progression rate, thus providing a basis for personalized diagnosis and treatment. Simultaneously, combined detection of genes related to muscular dystrophy that exhibit similar clinical manifestations to Duchenne muscular dystrophy can effectively improve differential diagnostic capabilities, reduce the risk of misdiagnosis and missed diagnosis, and simplify the traditional step-by-step testing process. Therefore, combining the detection of Duchenne muscular dystrophy pathogenic genes... DMD Integrating modified genes and differential diagnostic genes into the same detection system is of great significance for improving detection coverage, enhancing diagnostic accuracy, and achieving refined clinical management. Summary of the Invention

[0008] at present DMD The detection technologies used in clinical applications all have problems such as being unable to explain differences in patient clinical phenotypes, difficulty in prognostic assessment, and easy confusion with other hereditary myopathy. In the molecular diagnostic system for Duchenne muscular dystrophy, [the following will be addressed / addressed]: DMD The combined detection of modifying genes and genes related to differential diagnosis has become an important development direction in recent years. This strategy not only expands the diagnostic dimensions of traditional single-gene testing, but also has significant implications for clinical decision-making and prognostic assessment, demonstrating excellent technical effects in improving diagnostic accuracy, refining clinical triage, and reducing misdiagnosis and missed diagnosis.

[0009] In view of this, the present invention provides a detection method. DMD Gene, DMD Probe combinations, kits, and methods for modifying genes and identifying gene variations.

[0010] On one hand, the present invention provides a probe assembly comprising probes (1) and (6) of the following probe groups, which is also referred to herein as probe assembly A: (1) Targeting DMD probe sets of gene regions, wherein DMD Gene regions contain DMD Full-length gene; (2) Targeting DMD The probe set of gene regulatory regions, wherein DMD Gene regulatory regions include DMD The 20 kb region upstream of the 5'UTR and the 20 kb region downstream of the 3'UTR of the gene; (3) Targeting DMD The probe set that modifies the gene region, wherein DMD Modified gene regions include SPP1 Key gene loci rs28357094, rs11730582, rs11439060, and rs17524488. LTBP4 Key gene loci: rs2303729, rs1131620, rs1051303, rs710160, and rs10880. CD40 The key gene site rs1883832 ACTN3 The key gene locus rs1815739 DYNLT5 Key gene loci rs1060575 and rs3816989, THBS1 Key gene loci rs2725797 and rs2624259, and ADRB2 The key gene locus is rs1042713. (4) A probe set targeting a diagnostic gene region, wherein the diagnostic gene region comprises the full length of the following genes:LMNA, PABPN1, CAPN3, DMPK, DYSF, TTN, COL6A1, CNBP, LAMA2, GNE, EMD, SGCA, SGCG, SGCB, SGCD, DYNLT5, FKRP, ANO5, CRPPA, POMT2, FKTN, POMGNT1, POMT1, DAG1, GMPPB, POMK, STIM1 ; (5) A probe set targeting mitochondrial gene regions, wherein the mitochondrial gene regions contain the full length of the following genes: ND1, ND2, ND3, ND4, ND4L, ND5, ND6, CYTB, COX1, COX2, COX3, ATP6, ATP8, MT-RNR1, MT-RNR2, MT-TA (Ala), MT-TR (Arg), MT-TN (Asn), MT-TD (Asp), MT-TC (Cys), MT-TE (Glu), MT-TQ (Gln), MT-TG (Gly), MT-TH (His), MT-TI (Ile), MT-TL1, MT-TL2, MT-TK (Lys), MT-TM (Met), MT-TF (Phe), MT-TP (Pro), MT-TS1, MT-TS2, MT-TT (Thr), MT-TW (Trp), MT-TY (Tyr), MT-TV (Val); The aforementioned mitochondrial gene region contains 13 genes encoding polypeptides: ND1, ND2, ND3, ND4, ND4L, ND5, ND6, CYTB, COX1, COX2, COX3, ATP6, and ATP8; 2 rRNA genes: MT-RNR1 and MT-RNR2; and 22 tRNA genes: MT-TA (Ala), MT-TR (Arg), MT-TN (Asn), MT-TD (Asp), MT-TC (Cys), MT-TE (Glu), MT-TQ (Gln), MT-TG (Gly), MT-TH (His), MT-TI (Ile), MT-TL1, MT-TL2, MT-TK (Lys), MT-TM (Met), MT-TF (Phe), MT-TP (Pro), MT-TS1, MT-TS2, MT-TT (Thr), MT-TW (Trp), MT-TY (Tyr), and MT-TV (Val).

[0011] (6) A probe set targeting the sex identification gene region, wherein the sex identification gene region includes the full length of the SRY gene and its 2 kb upstream region of the 5'UTR and its 2 kb downstream region of the 3'UTR.

[0012] In summary, the detection sites of the probe combination described in this article include: DMD, SPP1, LTBP4, CD40, ACTN3, THBS1, ADRB2, LMNA, PABPN1, CAPN3, DMPK, DYSF, TTN, COL6A1, CNBP, LAMA2, GNE, EMD, SGCA, SGCG, SGCB, SGCD, DYNLT5, FKRP, ANO5, CRPPA, POMT2, FKTN, POMGNT1, POMT1, DAG1, GMPPB, POMK, STIM1 , SRY, ND1, ND2, ND3, ND4, ND4L, ND5, ND6, CYTB, COX1, COX2, COX3, ATP6, ATP8, MT- RNR1, MT-RNR2, MT-TA (Ala), MT-TR (Arg), MT-TN (Asn), MT-TD (Asp), MT-TC (Cys), MT-TE (Glu), MT-TQ (Gln), MT-TG (Gly), MT-TH (His), MT-TI (Ile), MT-TL1, MT-TL2, MT-TK (Lys), MT-TM (Met), MT-TF (Phe), MT-TP (Pro), MT-TS1, MT-TS2, MT-TT (Thr), MT-TW (Trp), MT-TY (Tyr), MT-TV (Val) The genes, the 3'UTR and 5'UTR regions of the aforementioned genes; and the key sites rs28357094, rs11730582, rs11439060, rs17524488, rs2303729, rs1131620, rs1051303, rs710160, rs10880, rs1883832, rs1815739, rs1060575, rs3816989, rs2725797, rs2624259 and rs1042713.

[0013] In some embodiments, the probe assembly comprises probe groups (1), (2), (3), and (6). This probe assembly is also referred to herein as probe assembly D.

[0014] In some embodiments, the probe assembly comprises probe groups (1), (2), (3), (4), (5), and (6). This probe assembly is also referred to herein as probe assembly B.

[0015] In some implementations, the above-mentioned sites are derived from humans, with the reference sequence being the human reference genome version GRCh38.

[0016] In some implementations, the DMD Gene regulatory regions include DMD The 100 kb region upstream of the 5'UTR and the 100 kb region downstream of the 3'UTR of the gene.

[0017] In some implementations, the DMD Modified gene regions further include SPP1 Full-length gene LTBP4 Full-length gene CD40 Full-length gene ACTN3 Full-length gene DYNLT5 Full-length gene THBS1 Full-length gene and ADRB2 Full-length gene.

[0018] In some implementations, the DMD Modified gene regions further include SPP1 Full-length gene and its 5'UTR and 3'UTR regions, LTBP4 Full-length gene and its 5'UTR and 3'UTR regions, CD40 Full-length gene and its 5'UTR and 3'UTR regions, ACTN3 Full-length gene and its 5'UTR and 3'UTR regions, DYNLT5 Full-length gene and its 5'UTR and 3'UTR regions, THBS1Full-length gene and its 5'UTR and 3'UTR regions and ADRB2 Full-length gene and its 5'UTR and 3'UTR regions.

[0019] In some implementations, each probe in the probe assemblies described herein meets the following requirement: (a) targeting PABPN1, CAPN3, DMPK (a) The full-length probe covers 1-3 kb regions of the 5'UTR and 3'UTR and is designed with a 1× flat coverage method; the remaining probes evenly cover the target region and the interval between each two adjacent probes is 400-1000 bp; (b) For regions where probe design is not suitable, such as those with poor sequence specificity, no probes are designed; (c) The probe length is 80-120 bp and the GC content is 40%-60%.

[0020] In some implementations, each probe in the probe ensemble described herein is a DNA probe or an RNA probe.

[0021] In some embodiments, each probe in the probe assemblies described herein is linked to a marker selected from at least one or more of biotin, avidin, streptavidin, antibodies, or chemical conjugates.

[0022] In some implementations, each probe group in the probe assemblies described herein includes probes covering one, more, or all of the following coverage types: (i) probes covering the entire length; (ii) probes covering exon regions; (iii) probes covering variant hotspot regions; and (iv) probes covering key SNP sites. The probe groups involved in this invention cover gene sites in four types as described above. Different probe coverage types can meet different clinical application scenarios, achieving the most effective detection with the most cost-efficient sequencing.

[0023] In some implementations, the probe combination described herein is as follows: Probe assembly (A), comprising probe set (1) and probe set (6), for the diagnosis of Duchenne muscular dystrophy; The probe set (B), which includes probe group (1), probe group (2), probe group (3), probe group (4), probe group (5) and probe group (6), is used for the diagnosis and differential diagnosis analysis of Duchenne muscular dystrophy. Probe set (C), comprising probe group (4), probe group (5), and probe group (6), is used for differential diagnostic analysis of Duchenne muscular dystrophy; or Probe assembly (D), comprising probe group (1), probe group (2), probe group (3), and probe group (6), is used for the diagnosis of Duchenne muscular dystrophy. DMD Analysis of regulatory and modifying genes.

[0024] In some specific implementation plans: (1) Targeting DMDSpecific probe sequences for the gene region probe set may include: chrX:31109561-31109661, chrX:31110058-31110158, chrX:31110558-31110658, chrX:31111058-31111158, chrX:31111922-31112022, chrX:31112423-31112523, chrX:31112933-31113033, chrX:31113433-31113533, chrX:31114002-31114102, chrX:31114552-311146 52, chrX:31115049-31115149, chrX:31115549-31115649, chrX:3111604 9-31116149,chrX:31116611-31116711,chrX:31117101-31117201,chrX: 31117611-31117711,chrX:31118109-31118209,chrX:31118221-311183 21, chrX:31118711-31118811, chrX:31119211-31119311, chrX:31119711 -31119811,chrX:31120277-31120377,chrX:31120777-31120877,chrX: 31121267-31121367,chrX:31121767-31121867,chrX:31122268-3112236 8. chrX:31122768-31122868, chrX:31123270-31123370, chrX:31123762 -31123862,chrX:31124262-31124362,chrX:31124753-31124853,chrX:3 1125253-31125353,chrX:31125753-31125853,chrX:31126253-3112635 3. chrX:31126823-31126923, chrX:31127315-31127415, 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83536-33283636,chrX:33284125-33284225,chrX:33284615-33284715,chrX:33285106-33285206,chrX:33285606-33285706,chrX:33286106-33286206,chrX:33286651-33286751,chrX:33287151-33287251,chrX:33287652-33287752,chrX:33288152-33288252,chrX:33288648-33288748,chrX:33289148-33289248,chrX:33289648-33289748,chrX:33290146-33290246,chrX:33290646-33290746,chrX:33291466-33291566,chrX:33291965-33292065,chrX:33292464-33292564,chrX:33292964-33293064,chrX:33293454-33293554,chrX:33293963-33294063,chrX:33294463-33294563,chrX:33294963-33295063,chrX:33295493-33295593,chrX:33295994-33296094,chrX:33296493-33296593,chrX:33296993-33297093,chrX:33297652-33297752,chrX:33298152-33298252,chrX:33298652-33298752,chrX:33299158-33299258,chrX:33299658-33299758,chrX:33300158-33300258,chrX:33300748-33300848,chrX:33301248-33301348,chrX:33301868-33301968,chrX:33302358-33302458,chrX:33302858-33302958,chrX:33303367-33303467,chrX:33303870-33303970,chrX:33305614-33305714,chrX:33306109-33306209,chrX:33306609-33306709,chrX:33307117-33307217,chrX:33307685-33307785,chrX:33308185-33308285,chrX:33308685-33308785,chrX:33309184-33309284,chrX:33309682-33309782,chrX:33310182-33310282,chrX:33310681-33310781,chrX:33311250-33311350,chrX:33311759-33311859,chrX:33312254-33312354,chrX:33312757-33312857,chrX:33313257-33313357,chrX:33313757-33313857,chrX:33314687-33314787,chrX:33315197-33315297,chrX:33315697-33315797,chrX:33316187-33316287,chrX:33316697-33316797,chrX:33317193-33317293,chrX:33317698-33317798,chrX:33318198-33318298,chrX:33318728-33318828,chrX:33319218-33319318,chrX:33319718-33319818,chrX:33320218-33320318,chrX:33320985-33321085,chrX:33322026-33322126,chrX:33322526-33322626,chrX:33323016-33323116,chrX:33323506-33323606,chrX:33324001-33324101,chrX:33324509-33324609,chrX:33325016-33325116,chrX:33325516-33325616,chrX:33326016-33326116,chrX:33326516-33326616,chrX:33327025-33327125,chrX:33327521-33327621,chrX:33328017-33328117,chrX:33328517-33328617,chrX:33329022-33329122,chrX:33329522-33329622,chrX:33330128-33330228,chrX:33330627-33330727,chrX:33331128-33331228,chrX:33331628-33331728,chrX:33332135-33332235,chrX:33332627-33332727,chrX:33333125-33333225,chrX:33333621-33333721,chrX:33334121-33334221,chrX:33334628-33334728,chrX:33335128-33335228,chrX:33335628-33335728,chrX:33336128-33336228,chrX:33336628-33336728,chrX:33337133-33337233,chrX:33337633-33337733,chrX:33338133-33338233,chrX:33338633-33338733,chrX:33339123-33339223,chrX:33339623-33339723;, (2) Specific probe sequences targeting the DMD gene regulatory region may include: chrX:31099221-31099321, chrX:31099721-31099821, chrX:31100221-31100321, chrX:31100721-31100821, chrX:31101546-31101646, chrX:31102151-31102251, chrX:31102651-31102751, chrX:31103151-31103251, chrX:31103644-31103744, chrX:3110413 8-31104238,chrX:31104828-31104928,chrX:31105378-31105478,chrX: 31105875-31105975,chrX:31107021-31107121,chrX:31107517-3110761 7. chrX:31108026-31108126, chrX:31108529-31108629, chrX:31109020- 31109120,chrX:33340131-33340231,chrX:33340388-33340488,chrX:333 40878-33340978, chrX:33341448-33341548, chrX:33342028-33342128, c hrX:33342550-33342650, chrX:33343156-33343256, chrX:33343802-333 43902, chrX:33344388-33344488, chrX:33344888-33344988, chrX:33345 418-33345518,chrX:33347428-33347528,chrX:33348269-33348369,chrX :33348773-33348873,chrX:33349273-33349373,chrX:33349773-333498 73, chrX:33350278-33350378, chrX:33350788-33350888, chrX:33351296 -33351396,chrX:33351796-33351896,chrX:33352296-33352396,chrX:3 3352886-33352986,chrX:33353386-33353486,chrX:33353886-33353986,chrX: 33354386-33354486, chrX: 33354896-33354996, chrX: 33355386-33355486, chrX: 33355895-33355995, chrX: 33356385-33356485, chrX: 33356883-33356983, chrX: 33357393-33357493, chrX: 33357892-33357992, chrX: 33358431-33358531, chrX: 33359061-33359161;, (3) targeting DMDSpecific probe sequences for the probe set modifying gene regions may include: chr1:66750458-66750558, chr1:66750955-66751055, chr1:66751455-66751555, chr1:66751955-66752055, chr1:66752455-66752555, chr1:66752950-66753050, chr1:66753450-66753550, chr1:66754236-66754336, chr1:66754736-66754836, chr1:66755236-6675 5336, chr1:66755731-66755831, chr1:66756231-66756331, chr1:66756 731-66756831,chr1:66757231-66757331,chr1:66757731-66757831,chr 1:66758231-66758331, chr1:66758722-66758822, chr1:66759222-66759 322, chr1:66759722-66759822, chr1:66760292-66760392, chr1:6676079 4-66760894,chr1:66761384-66761484,chr1:66762112-66762212,chr1 :66762633-66762733,chr1:66763313-66763413,chr1:66764480-667645 80, chr1:66764973-66765073, chr1:66765473-66765573, chr1:66765973 -66766073,chr1:66766473-66766573,chr1:66767118-66767218,chr1:6 6767609-66767709,chr1:66768108-66768208,chr1:66768602-6676870 2. chr1:66769102-66769202, chr1:66769608-66769708, chr1:66770108- 66770208,chr1:66770374-66770474,chr1:66770608-66770708,chr1:66 771108-66771208,chr1:66771608-66771708,chr1:66772198-66772298,chr1:66772703-66772803,chr1:66773203-66773303,chr1:66773703-66773803,chr1:66774194-66774294,chr1:66774694-66774794,chr1:66775194-66775294,chr1:66775694-66775794,chr1:66776194-66776294,chr1:66776398-66776498,chr1:66776694-66776794,chr1:66777194-66777294,chr1:66777690-66777790,chr1:66778190-66778290,chr1:66778687-66778787,chr1:66779187-66779287,chr1:66779687-66779787,chr11:66544399-66544499,chr11:66545322-66545422,chr11:66545822-66545922,chr11:66546322-66546422,chr11:66546822-66546922,chr11:66547322-66547422,chr11:66547815-66547915,chr11:66548315-66548415,chr11:66548815-66548915,chr11:66549315-66549415,chr11:66549815-66549915,chr11:66550311-66550411,chr11:66550837-66550937,chr11:66551337-66551437,chr11:66552378-66552478,chr11:66553896-66553996,chr11:66554476-66554576,chr11:66555026-66555126,chr11:66555523-66555623,chr11:66556028-66556128,chr11:66557048-66557148,chr11:66557540-66557640,chr11:66558031-66558131,chr11:66558651-66558751,chr11:66559145-66559245,chr11:66559645-66559745,chr11:66560145-66560245,chr11:66560577-66560745,chr11:6656114 5-66561245,chr11:66561637-66561737,chr11:66562128-66562228,chr 11:66562628-66562728,chr11:66563128-66563228,chr11:66563628-6 6563728,chr11:66564128-66564228,chr11:66564625-66564725,chr11: 66565125-66565225, chr15:38814777-38814877, chr15:38815345-38815445, chr15:38815701-38815801, chr15:38815845-38815945, chr15:388 16241-38816341, chr15:38816345-38816445, chr15:38816791-38816945, chr15:38816992-38817092, chr15:38817291-38817445, chr15:388177 97-38817945, chr15:38818342-38818445, chr15:38818662-38818762, chr15:38818842-38818945, chr15:38819342-38819445, chr15:38819842- 38819945, chr15:38820408-38820508, chr15:38821028-38821128, chr15:38821528-38821628, chr15:39579078-39579178, chr15:39579628-395 79728, chr15:39580128-39580228, chr15:39580628-39580728, chr15:39581124-39581224, chr15:39581659-39581759, chr15:39582159-395822 59, chr15:39582659-39582759, chr15:39583159-39583259, chr15:39583659-39583759, chr15:39584159-39584259, chr15:39584655-39584755chr15:39585154-39585254, chr15:39585654-39585754, chr15:39586240-39586340, chr15:39586740-39586840, chr15:39587240-39587340, chr 15:39587740-39587840, chr15:39588250-39588350, chr15:39588750-39588850, chr15:39589250-39589350, chr15:39589750-39589850, chr15: 39590250-39590350, chr15:39590750-39590850, chr15:39591250-39591350, chr15:39591750-39591850, chr15:39592240-39592340, chr15:395 92740-39592840, chr15:39593240-39593340, chr15:39593740-39593840, chr15:39594244-39594344, chr15:39594750-39594850, chr15:395952 40-39595340, chr15:39595740-39595840, chr15:39596243-39596343, chr15:39596736-39596836, chr15:39597297-39597397, chr15:39597797- 39597897, chr15:39598297-39598397, chr15:39598797-39598897, chr15:39599297-39599397, chr15:39599797-39599897, chr15:39600304-396 00404,chr15:39600806-39600906,chr15:39601306-39601406,chr19:4 0591165-40591265,chr19:40591805-40591905,chr19:40592298-405923 98,chr19:40592798-40592898,chr19:40593528-40593628,chr19:4059 4132-40594232,chr19:40594632-40594732,chr19:40595132-40595232,chr19:40595623-40595723, chr19:40596327-40596427, chr19:40596827-40596927, chr19:40597327-40597427, chr19:40597827-40597927, chr 19:40598327-40598427, chr19:40598896-40598996, chr19:40599394-40599494, chr19:40599894-40599994, chr19:40600394-40600494, chr19: 40600894-40600994, chr19:40601394-40601494, chr19:40601894-40601994, chr19:40602394-40602494, chr19:40602894-40602994, chr19:406 03394-40603494, chr19:40603894-40603994, chr19:40604394-40604494, chr19:40604894-40604994, chr19:40605142-40605242, chr19:406053 94-40605494, chr19:40605894-40605994, chr19:40606394-40606494, chr19:40607037-40607137, chr19:40607540-40607640, chr19:40608040- 40608140, chr19:40608540-40608640, chr19:40609030-40609130, chr19:40609530-40609630, chr19:40610030-40610130, chr19:40610530-406 10630, chr19:40611030-40611130, chr19:40611530-40611630, chr19:40611905-40612005, chr19:40612030-40612130, chr19:40612147-406122 47, chr19:40612530-40612630, chr19:40613030-40613130, chr19:40613530-40613630, chr19:40614030-40614130, chr19:40614530-40614630chr19:40615030-40615130, chr19:40615758-40615858, chr19:40616726-40616826, chr19:40617230-40617330, chr19:40617739-40617839, chr 19:40618659-40618759, chr19:40619159-40619259, chr19:40619659-40619759, chr19:40620149-40620249, chr19:40621230-40621330, chr19: 40622040-40622140, chr19:40622364-40622464, chr19:40622532-40622632, chr19:40623028-40623128, chr19:40623528-40623628, chr19:406 24028-40624128, chr19:40624738-40624838, chr19:40625408-40625508, chr19:40625908-40626008, chr19:40626415-40626515, chr19:406269 15-40627015, chr19:40627416-40627516, chr19:40627916-40628016, chr19:40628516-40628616, chr19:40629135-40629235, chr19:40629644- 40629744, chr19:40630534-40630634, chr19:40631184-40631284, chr20:46116313-46116413, chr20:46116863-46116963, chr20:46117632-461 17732, chr20:46118132-46118232, chr20:46118303-46118403, chr20:46118632-46118732, chr20:46119132-46119232, chr20:46119631-461197 31, chr20:46120128-46120228, chr20:46120628-46120728, chr20:46121126-46121226, chr20:46121626-46121726, chr20:46122126-46122226chr20:46122618-46122718,chr20:46123121-46123221,chr20:46123621-46123721,chr20:46124331-46124431,chr20:46125056-46125156,chr20:46125562-46125662,chr20:46126062-46126162,chr20:46126610-46126710,chr20:46127139-46127239,chr20:46127648-46127748,chr20:46128146-46128246,chr20:46128647-46128747,chr20:46129147-46129247,chr20:46129646-46129746,chr20:46130146-46130246,chr20:46130637-46130737,chr20:46131645-46131745,chr4:87973713-87973813,chr4:87974213-87974313,chr4:87974713-87974813,chr4:87975203-87975313,chr4:87975554-87975705,chr4:87975714-87975814,chr4:87976214-87976314,chr4:87976713-87976813,chr4:87977212-87977312,chr4:87977712-87977812,chr4:87978220-87978320,chr4:87978720-87978820,chr4:87979460-87979560,chr4:87979959-87980059,chr4:87980459-87980559,chr4:87980959-87981059,chr4:87981451-87981551,chr4:87981947-87982047,chr4:87982457-87982557,chr4:87982958-87983058,chr4:87983468-87983568,chr4:87984128-87984228,chr4:87984618-87984718,chr4:87985118-87985218,chr5:148824610-148824710,chr5:148825352-148825452, chr5:148825852-148825952, chr5:148826358-148826458, chr5:148826837-148826958, chr5:148827358-148827458, chr5:14882786 0-148827960, chr5:148828360-148828460, chr5:148828860-148828960, chr5:148829410-148829510, chr5:148829910-148830010, chr5:148830417-148830517; (4) Specific probe sequences for the probe set targeting the differential diagnostic gene region may include: chr1:156080572-156080672, chr1:156081072-156081172, chr1:156081576-156081676, chr1:156082080-156082180, chr1:156082580-156082680, chr1:156083080-156083180, chr1:156083807-156083907, chr1:156084307-156084407, chr1:156084807-1 56084907,chr1:156085299-156085399,chr1:156086061-156086161,ch r1:156086551-156086651, chr1:156087061-156087161, chr1:15608815 0-156088250,chr1:156089620-156089720,chr1:156090124-156090224 , chr1:156090634-156090734, chr1:156091135-156091235, chr1:156091 637-156091737,chr1:156092686-156092786,chr1:156093186-1560932 86, chr1:156093679-156093779, chr1:156094169-156094269, chr1:156 094669-156094769,chr1:156095217-156095317,chr1:156095717-1560 95817,chr1:156096225-156096325,chr1:156096725-156096825,chr1: 156097233-156097333,chr1:156097733-156097833,chr1:156098233-1 56098333,chr1:156098731-156098831,chr1:156099237-156099337,ch r1:156099879-156099979, chr1:156100379-156100479, chr1:15610088 0-156100980,chr1:156101520-156101620,chr1:156102026-156102126,chr1:156102523-156102623,chr1:156103023-156103123,chr1:156103523-156103623,chr1:156104030-156104130,chr1:156104530-156104630,chr1:156105020-156105120,chr1:156105529-156105629,chr1:156106139-156106239,chr1:156106648-156106748,chr1:156107148-156107248,chr1:156107648-156107748,chr1:156108148-156108248,chr1:156108774-156108874,chr1:156109272-156109372,chr1:156110134-156110234,chr1:156110964-156111064,chr1:156111464-156111564,chr1:156111960-156112060,chr1:156112460-156112560,chr1:156113327-156113427,chr1:156113828-156113928,chr1:156114326-156114426,chr1:156114826-156114926,chr1:156115326-156115426,chr1:156115826-156115926,chr1:156116336-156116436,chr1:156117745-156117845,chr1:156118245-156118345,chr1:156118745-156118845,chr1:156119541-156119641,chr1:156120041-156120141,chr1:156120724-156120824,chr1:156121313-156121413,chr1:156122123-156122223,chr1:156122622-156122722,chr1:156123122-156123222,chr1:156123623-156123723,chr1:156124123-156124223,chr1:156124623-156124723,chr1:156125124-156125224,chr1:156125994-156126094,chr1:156126494-156126594,chr1:156126994-156127094,chr1:156127979-156128079,chr1:156128471-156128571,chr1:156128969-156129069,chr1:156129469-156129569,chr1:156129964-156130064,chr1:156130464-156130564,chr1:156131604-156131704,chr1:156132494-156132594,chr1:156133584-156133684,chr1:156134187-156134287,chr1:156134687-156134787,chr1:156135188-156135288,chr1:156135692-156135792,chr1:156136192-156136292,chr1:156136692-156136792,chr1:156137197-156137297,chr1:156137697-156137797,chr1:156138197-156138297,chr1:156138697-156138797,chr1:156139197-156139297,chr1:156139692-156139792,chr1:156140192-156140292,chr1:156140692-156140792,chr1:156141192-156141292,chr14:23319462-23320248,chr14:23320251-23321429,chr14:23321507-23321607,chr14:23321616-23322200,chr14:23322209-23323383,chr14:23323392-23323588,chr15:42357500-42357600,chr15:42358000-42358100,chr15:42358500-42358600,chr15:42359000-42359100,chr15:42359505-42359605,chr15:42360005-42360105,chr15:42360505-42360605,chr15:42361005-42361105, chr15:42361505-42361605, chr15:42362005-42362105, chr15:42362505-42362605, chr15:42363015-42363115, chr 15:42363523-42363623, chr15:42364023-42364123, chr15:42364523-42364623, chr15:42365023-42365123, chr15:42365523-42365623, chr15: 42366023-42366123, chr15:42366523-42366623, chr15:42367103-42367203, chr15:42367603-42367703, chr15:42368103-42368203, chr15:423 68631-42368731, chr15:42369501-42369601, chr15:42370151-42370251, chr15:42370656-42370756, chr15:42371156-42371256, chr15:423720 73-42372173, chr15:42372827-42372927, chr15:42373335-42373435, chr15:42373835-42373935, chr15:42374337-42374437, chr15:42375123- 42375223, chr15:42375613-42375713, chr15:42376103-42376203, chr15:42376633-42376733, chr15:42377180-42377280, chr15:42377690-423 77790, chr15:42378320-42378420, chr15:42378830-42378930, chr15:42379340-42379440, chr15:42380108-42380208, chr15:42380636-423807 36, chr15:42381129-42381229, chr15:42381819-42381919, chr15:42382647-42382747, chr15:42383142-42383242, chr15:42383866-42383966chr15:42384435-42384535, chr15:42384944-42385044, chr15:42385553-42385653, chr15:42386053-42386153, chr15:42386552-42386652, chr 15:42387052-42387152, chr15:42387552-42387652, chr15:42388054-42388154, chr15:42388561-42388661, chr15:42389061-42389161, chr15: 42389561-42389661, chr15:42390061-42390161, chr15:42390561-42390661, chr15:42391084-42391184, chr15:42391582-42391682, chr15:423 92187-42392287, chr15:42392695-42392795, chr15:42393197-42393297, chr15:42393884-42393984, chr15:42394384-42394484, chr15:423948 84-42394984, chr15:42395384-42395484, chr15:42395884-42395984, chr15:42396514-42396614, chr15:42397014-42397114, chr15:42397654- 42397754, chr15:42399224-42399324, chr15:42399724-42399824, chr15:42400224-42400324, chr15:42400724-42400824, chr15:42401217-424 01317, chr15:42401722-42401822, chr15:42402222-42402322, chr15:42402722-42402822, chr15:42403225-42403325, chr15:42403725-424038 25, chr15:42404225-42404325, chr15:42404725-42404825, chr15:42405215-42405315, chr15:42405715-42405815, chr15:42406261-42406361chr15:42406757-42406857,chr15:42407247-42407347,chr15:4240774 7-42407847,chr15:42408247-42408347,chr15:42408747-42408847,chr 15:42409239-42409339,chr15:42409739-42409839,chr15:42410239-4 2410339,chr15:42410739-42410839,chr15:42411239-42411339,chr15: 42411739-42411839,chr15:42412239-42412339,chr15:42412740-4241 2840,chr15:42413240-42413340,chr15:42413747-42413847,chr15:424 14249-42414349, chr19:40591795-40591895, chr19:40592294-40592394, chr19:40592794-40592894, chr19:40593532-40593632, chr19:405941 22-40594222, chr19:40594621-40594721, chr19:40595121-40595221, chr19:40595621-40595721, chr19:40607034-40607134, chr19:40617226- 40617326, chr19:40617736-40617836, chr19:40620836-40620936, chr19:40621466-40621566, chr19:40622032-40622132, chr19:40622534-406 22634, chr19:40629635-40629735, chr19:40630536-40630636, chr19:40631166-40631266, chr19:45768195-45768595, chr19:45768597-457686 97, chr19:45768704-45768804, chr19:45768805-45770196, chr19:45770246-45770729, chr19:45770739-45770839, 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chr6:129129933-129130033, chr6:129130933-129131033, chr6:129131923-129132023, chr6:129132913-129133013, chr6:129133911-129134011, chr6:129134901-129135001, chr6:129135911-129136011, chr6:129136908-129137008, chr6:129137908-129138008, chr6:129138899-129138999, chr6:129139899-129139999, chr6:129140899-129140999, chr6:129141899-129141999, chr6:129142899-129142999,chr6:129143904-129144004, chr6:129144904-129145004, chr6:129145904-129146004, chr6:129146903-129147003, chr6:129148026-129148126, chr6:129149023-129149123, chr6:129150022-129150122, chr6:129151022-129151122, chr6:129152012-129152112, chr6:129153012-129153112, chr6:129154002-129154102, chr6:129154993-129155093, chr6:129156173-129156273, chr6:129157182-129157282, chr6:129158182-129158282, chr6:129159184-129159284, chr6:129160187-129160287, chr6:129161185-129161285, chr6:129162194-129162294, chr6:129163199-129163299, chr6:129164199-129164299, chr6:129165198-129165298, chr6:129166196-129166296, chr6:129173166-129173266, chr6:129174157-129174257, chr6:129175154-129175254, chr6:129176151-129176251, chr6:129177143-129177243, chr6:129178143-129178243, chr6:129179143-129179243, chr6:129180142-129180242, chr6:129181132-129181232, chr6:129182132-129182232, chr6:129183128-129183228, chr6:129184138-129184238, chr6:129185138-129185238, chr6:129186224-129186324, chr6:129187224-129187324, chr6:129188224-129188324, chr6:129189224-129189324,chr6:129190224-129190324, chr6:129191215-129191315, chr6:129192212-129192312, chr6:129193212-129193312, chr6:129194212-129194312, chr6:129195211-129195311, chr6:129196211-129196311, chr6:129197221-129197321, chr6:129198231-129198331, chr6:129199261-129199361, chr6:129200546-129200646, chr6:129201546-129201646, chr6:129202536-129202636, chr6:129203541-129203641, chr6:129204531-129204631, chr6:129205532-129205632, chr6:129206532-129206632, chr6:129207522-129207622, ​​chr6:129208748-129208848, chr6:129210018-129210118, chr6:129211014-129211114, chr6:129212004-129212104, 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chr6:129335166-129335266, chr6:129336292-129336392, chr6:129337292-129337392, chr6:129338292-129338392, chr6:129339292-129339392, chr6:129340298-129340398, chr6:129341298-129341398, chr6:129342306-129342406, chr6:129343306-129343406, chr6:129344306-129344406, chr6:129345311-129345411, chr6:129346311-129346411, chr6:129347311-129347411, chr6:129348321-129348421, chr6:129349321-129349421, chr6:129350321-129350421, chr6:129351321-129351421, chr6:129352321-129352421, chr6:129353321-129353421, chr6:129354331-129354431, chr6:129355330-129355430, chr6:129356330-129356430, chr6:129357339-129357439, chr6:129358339-129358439,chr6:129359336-129359436, chr6:129360336-129360436, chr6:129361332-129361432, chr6:129362324-129362424, chr6:129363314-129363414, chr6:129364319-129364419, chr6:129365688-129365788, chr6:129366698-129366798, chr6:129367708-129367808, chr6:129368708-129368808, chr6:129369706-129369806, chr6:129370706-129370806, chr6:129371893-129371993, chr6:129372943-129373043, 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chr6:129413752-129413852, chr6:129414752-129414852, chr6:129415834-129415934, chr6:129416838-129416938, chr6:129417839-129417939, chr6:129419020-129419120, chr6:129420020-129420120, chr6:129421025-129421125, chr6:129422025-129422125, chr6:129423024-129423124, chr6:129424024-129424124, chr6:129425024-129425124, chr6:129426024-129426124, chr6:129427029-129427129, chr6:129428029-129428129, chr6:129429032-129429132, chr6:129430032-129430132, chr6:129431027-129431127, chr6:129432031-129432131, chr6:129433035-129433135, chr6:129434035-129434135, chr6:129435035-129435135, chr6:129436027-129436127, chr6:129437037-129437137, chr6:129438037-129438137, chr6:129439197-129439297, chr6:129440197-129440297, chr6:129441199-129441299,chr6:129442199-129442299, chr6:129443199-129443299, chr6:129444199-129444299, chr6:129445199-129445299, chr6:129446199-129446299, chr6:129447190-129447290, chr6:129448250-129448350, chr6:129449313-129449413, chr6:129450603-129450703, chr6:129451603-129451703, chr6:129452611-129452711, chr6:129453609-129453709, chr6:129454611-129454711, chr6:129455614-129455714, chr6:129456624-129456724, chr6:129457620-129457720, chr6:129458619-129458719, chr6:129459619-129459719, chr6:129460620-129460720, chr6:129461621-129461721, chr6:129462631-129462731, chr6:129463632-129463732, chr6:129464632-129464732, chr6:129465632-129465732, chr6:129466642-129466742, chr6:129467642-129467742, chr6:129468642-129468742, chr6:129469635-129469735, chr6:129470630-129470730, chr6:129471620-129471720, chr6:129472620-129472720, chr6:129473620-129473720, chr6:129474618-129474718, chr6:129475618-129475718, chr6:129476618-129476718, chr6:129477618-129477718, chr6:129478608-129478708, chr6:129479618-129479718, chr6:129480618-129480718, chr6:129481618-129481718,chr6:129482618-129482718, chr6:129483628-129483728, chr6:129484638-129484738, chr6:129485645-129485745, chr6:129486645-129486745, chr6:129487705-129487805, chr6:129488705-129488805, chr6:129489705-129489805, chr6:129490705-129490805, chr6:129491705-1294918 05, chr6:129492709-129492809, chr6:129493709-129493809, chr6:129494709-129494809, chr6:129495709-129495809, chr6:129496713-129496813, chr6:129497713-129497813, chr6:129498713-129498813, chr6:129500000-129500100, chr6:129501000-129501100, chr6:129501993-1295 02093, chr6:129502993-129503093, chr6:129503993-129504093, chr6:129505003-129505103, chr6:129506373-129506473, chr6:129507382-129507482, chr6:129508412-129508512, chr6:129509406-129509506, chr6:129510413-129510513, chr6:129511417-129511517, chr6:129512417-1 29512517, chr6:129513417-129513517, chr6:129514417-129514517, chr6:129515417-129515517, chr6:129516427-129516527, chr9:36212440-36212540, chr9:36212940-36213040, chr9:36213440-36213540, chr9:36214270-36214370, chr9:36214770-36214870, chr9:36215300-36215400,chr9:36215795-36215895, chr9:36216961-36217061, chr9:36217459-36217559, chr9:36217959-36218059, chr9:36218453-36218553, chr9:362 18953-36219053, chr9:36219455-36219555, chr9:36219955-36220055, chr9:36220455-36220555, chr9:36220951-36221051, chr9:36221461-362 21561, chr9:36221961-36222061, chr9:36222459-36222559, chr9:36222959-36223059, chr9:36223459-36223559, chr9:36224039-36224139, ch r9:36224869-36224969, chr9:36225779-36225879, chr9:36226468-36226568, chr9:36226968-36227068, chr9:36227471-36227571, chr9:362281 60-36228260, chr9:36228810-36228910, chr9:36229306-36229406, chr9:36229814-36229914, chr9:36230313-36230413, chr9:36230883-36230 983, chr9:36231464-36231564, chr9:36231964-36232064, chr9:36232458-36232558, chr9:36232950-36233050, chr9:36233672-36233772, chr9: 36234166-36234266, chr9:36234659-36234759, chr9:36235181-36235281, chr9:36235677-36235777, chr9:36236485-36236585, chr9:36236984- 36237084, chr9:36237484-36237584, chr9:36241394-36241494, chr9:36242164-36242264, chr9:36243329-36243429, chr9:36243827-36243927chr9:36244323-36244423, chr9:36245271-36245371, chr9:36245773-36245873, chr9:36246273-36246373, chr9:36246938-36247038, chr9:362 47438-36247538, chr9:36248039-36248139, chr9:36248609-36248709, chr9:36249109-36249209, chr9:36249890-36249990, chr9:36250391-362 50491, chr9:36250957-36251057, chr9:36251565-36251665, chr9:36252267-36252367, chr9:36252767-36252867, chr9:36253568-36253668, ch r9:36254518-36254618, chr9:36255018-36255118, chr9:36255628-36255728, chr9:36256538-36256638, chr9:36257121-36257221, chr9:362578 30-36257930, chr9:36258329-36258429, chr9:36258828-36258928, chr9:36259328-36259428, chr9:36259948-36260048, chr9:36260452-36260 552, chr9:36260943-36261043, chr9:36261556-36261656, chr9:36262056-36262156, chr9:36262556-36262656, chr9:36263056-36263156, chr9: 36263564-36263664, chr9:36264430-36264530, chr9:36264928-36265028, chr9:36265420-36265520, chr9:36266282-36266382, chr9:36267063- 36267163, chr9:36267712-36267812, chr9:36268217-36268317, chr9:36268717-36268817, chr9:36269227-36269327, 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264331-22264431,chr11:22264833-22264933,chr11:22265332-22265432,chr11:22265897-22265997,chr11:22266396-22266496,chr11:22266900-22267000,chr11:22267400-22267500,chr11:22268150-22268250,chr11:22268650-22268750,chr11:22269096-22269196,chr11:22269613-22269713,chr11:22270113-22270213,chr11:22270635-22270735,chr11:22271354-22271454,chr11:22271760-22271860,chr11:22272339-22272439,chr11:22272839-22272939,chr11:22273410-22273510,chr11:22273921-22274021,chr11:22274405-22274505,chr11:22274836-22274936,chr11:22275417-22275517,chr11:22275891-22275991,chr11:22276452-22276552,chr11:22276955-22277055,chr11:22277455-22277555,chr11:22277914-22278014,chr11:22278393-22278493,chr11:22278993-22279093,chr11:22279527-22279627,chr11:22279972-22280072,chr11:22280472-22280572,chr11:22280897-22280997,chr11:22281357-22281457,chr11:22281796-22281896,chr11:22282307-22282407,chr11:22282751-22282851,chr11:22283254-22283354,chr11:22283699-22283799,chr11:22284282-22284382,chr11:22284694-22284794,chr13:23158507-23158607,chr13:23159024-23159124,chr13:23159544-23159644,chr13:23160044-23160144,chr13:23160557-23160657,chr13:23161004-23161104,chr13:23161429-23161529,chr13:23161861-23161961,chr13:23162861-23162961,chr13:23163371-23163471,chr13:23163868-23163968,chr13:23164283-23164383,chr13:23164782-23164882,chr13:23165316-23165416,chr13:23165909-23166009,chr13:23166509-23166609,chr13:23166972-23167072,chr13:23167389-23167489,chr13:23168357-23168457,chr13:23168857-23168957,chr13:23169257-23169357,chr13:23169804-23169904,chr13:23170394-23170494,chr13:23170835-23170935,chr13:23171302-23171402,chr13:23171901-23172001,chr13:23172435-23172535,chr13:23172884-23172984,chr13:23173384-23173484,chr13:23173901-23174001,chr13:23174401-23174501,chr13:23174901-23175001,chr13:23175417-23175517,chr13:23175956-23176056,chr13:23176525-23176625,chr13:23176956-23177056,chr13:23177467-23177567,chr13:2317798 8-23178088,chr13:23178502-23178602,chr13:23179097-23179197,chr 13:23179681-23179781, chr13:23180090-23180190, chr13:23180591-23180691, chr13:23181091-23181191, chr13:23181667-23181767, chr13: 23182204-23182304, Chr13:23182722-23182822, Chr13:23183231-23183331, Chr13:23184028-23184128, Chr13:23184577-23184677, Chr13:231 85149-23185249, chr13:23185649-23185749, chr13:23186110-23186210, chr13:23186510-23186610, chr13:23186962-23187062, chr13:231874 77-23187577, chr13:23187977-23188077, chr13:23188726-23188826, chr13:23189126-23189226, chr13:23189641-23189741, chr13:23190231- 23190331, Chr13:23190731-23190831, Chr13:23191259-23191359, Chr13:23191771-23191871, Chr13:23192245-23192345, Chr13:23192756-231 92856, Chr13:23193256-23193356, Chr13:23193775-23193875, Chr13:23194275-23194375, Chr13:23194732-23194832, Chr13:23195224-231953 24, Chr13:23195689-23195789, Chr13:23196189-23196289, Chr13:23196712-23196812, Chr13:23197267-23197367, Chr13:23197715-23197815chr13:23198199-23198299,chr13:23199113-23199213,chr13:2319959 7-23199697,chr13:23200036-23200136,chr13:23200514-23200614,chr 13:23201014-23201114, chr13:23201424-23201524, chr13:23201903-23202003, chr13:23202379-23202479, chr13:23202844-23202944, chr13: 23203284-23203384, Chr13:23203784-23203884, Chr13:23204308-23204408, Chr13:23204809-23204909, Chr13:23205324-23205424, Chr13:232 05773-23205873, chr13:23206245-23206345, chr13:23206811-23206911, chr13:23207326-23207426, chr13:23207926-23208026, chr13:232084 89-23208589, chr13:23208998-23209098, chr13:23209557-23209657, chr13:23210085-23210185, chr13:23210685-23210785, chr13:23211120- 23211220, Chr13:23211663-23211763, Chr13:23212263-23212363, Chr13:23212678-23212778, Chr13:23213079-23213179, Chr13:23213677-232 13777, Chr13:23214141-23214241, Chr13:23214639-23214739, Chr13:23215139-23215239, Chr13:23215681-23215781, Chr13:23216181-232162 81, Chr13:23216708-23216808, Chr13:23217208-23217308, Chr13:23217711-23217811, Chr13:23218213-23218313, Chr13:23218617-23218717chr13:23219371-23219471,chr13:23220489-23220589,chr13:2322100 4-23221104,chr13:23221469-23221569,chr13:23221959-23222059,chr 13:23222454-23222554, chr13:23222907-23223007, chr13:23223316-23223416, chr13:23224092-23224192, chr13:23224681-23224781, chr13: 23225277-23225377, Chr13:23225829-23225929, Chr13:23226329-23226429, Chr13:23226905-23227005, Chr13:23227440-23227540, Chr13:232 28034-23228134, chr13:23228496-23228596, chr13:23229304-23229404, chr13:23229813-23229913, chr13:23230624-23230724, chr13:232310 62-23231162, chr13:23231562-23231662, chr13:23232126-23232226, chr13:23232825-23232925, chr13:23233378-23233478, chr13:23233976- 23234076, Chr13:23234438-23234538, Chr13:23234915-23235015, Chr13:23235415-23235515, Chr13:23235871-23235971, Chr13:23236280-232 36380, Chr13:23236858-23236958, Chr13:23237378-23237478, Chr13:23237828-23237928, Chr13:23238231-23238331, Chr13:23238645-232387 45, Chr13:23239231-23239331, Chr13:23239726-23239826, Chr13:23240222-23240322, Chr13:23240626-23240726, Chr13:23241160-23241260chr13:23241565-23241665,chr13:23242163-23242263,chr13:2324264 5-23242745,chr13:23243131-23243231,chr13:23243631-23243731,chr 13:23244160-23244260, chr13:23244663-23244763, chr13:23245132-23245232, chr13:23245620-23245720, chr13:23246211-23246311, chr13: 23246684-23246784, Chr13:23247190-23247290, Chr13:23248119-23248219, Chr13:23249102-23249202, Chr13:23249667-23249767, Chr13:232 50267-23250367, chr13:23250678-23250778, chr13:23251178-23251278, chr13:23251734-23251834, chr13:23252320-23252420, chr13:232528 72-23252972, chr13:23253383-23253483, chr13:23253839-23253939, chr13:23254301-23254401, chr13:23254708-23254808, chr13:23255196- 23255296, chr13:23255739-23255839, chr13:23256206-23256306, chr13:23257371-23257471, chr13:23260849-23260949, chr13:23261409-232 61509, Chr13:23261951-23262051, Chr13:23262416-23262516, Chr13:23262899-23262999, Chr13:23263477-23263577, Chr13:23263897-232639 97, chr13:23264419-23264519, chr13:23264972-23265072, chr13:23265757-23265857, chr13:23266302-23266402, chr13:23266902-23267002chr13:23267328-23267428,chr13:23267828-23267928,chr13:2326835 7-23268457,chr13:23268807-23268907,chr13:23269308-23269408,chr 13:23269742-23269842, chr13:23270226-23270326, chr13:23270682-23270782, chr13:23271214-23271314, chr13:23271811-23271911, chr13: 23272274-23272374, Chr13:23272751-23272851, Chr13:23273153-23273253, Chr13:23273675-23273775, Chr13:23274202-23274302, Chr13:232 74769-23274869, chr13:23275530-23275630, chr13:23276004-23276104, chr13:23276724-23276824, chr13:23277134-23277234, chr13:232775 34-23277634, chr13:23278464-23278564, chr13:23278995-23279095, chr13:23279398-23279498, chr13:23279998-23280098, chr13:23280423- 23280523, Chr13:23280836-23280936, Chr13:23281405-23281505, Chr13:23281982-23282082, Chr13:23282738-23282838, Chr13:23284935-232 85035, chr13:23285343-23285443, chr13:23285786-23285886, chr13:23286286-23286386, chr13:23286784-23286884, chr13:23287284-232873 84, Chr13:23288060-23288160, Chr13:23288561-23288661, Chr13:23289077-23289177, Chr13:23289577-23289677, Chr13:23289997-23290097chr13:23290453-23290553,chr13:23290953-23291053,chr13:2329151 7-23291617,chr13:23291950-23292050,chr13:23292539-23292639,chr 13:23293039-23293139, chr13:23293460-23293560, chr13:23293968-23294068, chr13:23294483-23294583, chr13:23294983-23295083, chr13: 23295484-23295584, Chr13:23295980-23296080, Chr13:23296525-23296625, Chr13:23297012-23297112, Chr13:23297437-23297537, Chr13:232 98429-23298529, chr13:23298842-23298942, chr13:23299318-23299418, chr13:23299747-23299847, chr13:23300247-23300347, chr13:233007 19-23300819, chr13:23301132-23301232, chr13:23301532-23301632, chr13:23302093-23302193, chr13:23302670-23302770, chr13:23303224- 23303324, chr13:23303778-23303878, chr13:23304278-23304378, chr13:23305004-23305104, chr13:23305503-23305603, chr13:23306209-233 06309, chr13:23306780-23306880, chr13:23307320-23307420, chr13:23307845-23307945, chr13:23308398-23308498, chr13:23308986-233090 86, Chr13:23309483-23309583, Chr13:23309981-23310081, Chr13:23310478-23310578, Chr13:23311031-23311131, Chr13:23311593-23311693chr13:23312082-23312182,chr13:23312531-23312631,chr13:23312994-23313094,chr13:23313486-23313586,chr13:23314266-23314366,chr13:23314936-23315036,chr13:23315336-23315436,chr13:23315924-23316024,chr13:23317323-23317423,chr13:23317952-23318052,chr13:23318502-23318602,chr13:23318929-23319029,chr13:23319469-23319569,chr13:23319869-23319969,chr13:23320410-23320510,chr13:23320930-23321030,chr13:23321520-23321620,chr13:23322033-23322133,chr13:23322533-23322633,chr13:23323005-23323105,chr13:23323488-23323588,chr13:23324083-23324183,chr13:23324583-23324683,chr13:23325067-23325167,chr13:23325567-23325667,chr13:23326067-23326167,chr13:23326580-23326680,chr13:23327091-23327191,chr14:77272952-77273052,chr14:77273399-77273499,chr14:77273917-77274017,chr14:77274471-77274571,chr14:77275008-77275108,chr14:77275498-77275598,chr14:77275998-77276098,chr14:77276451-77276551,chr14:77276966-77277066,chr14:77277430-77277530,chr14:77277963-77278063,chr14:77278463-77278563,chr14:77278963-77279063,chr14:77279400-77279500, chr14:77279900-77280000, chr14:77280366-77280466, chr14:77281131-77281231, chr14:77281597-77281697, chr14:77282087-77282187, chr14:77282561-77282661, chr14:77283061-77283161, chr14:77283472-77283572, chr14:77283990-77284090, chr14: 77284490-77284590, chr14:77284938-77285038, chr14:77285438-77285538, chr14:77285943-77286043, chr14:77286541-77286641, chr14:77287126-77287226, chr14:77287683-77287783, chr14:77288225-77288325, chr14:77288725-77288825, chr14:77289467-77289567, chr14:772900 00-77290100, chr14:77290415-77290515, chr14:77290862-77290962, chr14:77291299-77291399, chr14:77292067-77292167, chr14:77292550-77292650, chr14:77293083-77293183, chr14:77293681-77293781, chr14:77294105-77294205, chr14:77294863-77294963, chr14:77295263-772 95363, chr14:77295763-77295863, chr14:77296163-77296263, chr14:77296753-77296853, chr14:77297243-77297343, chr14:77297743-77297843, chr14:77298243-77298343, chr14:77298784-77298884, chr14:77299312-77299412, chr14:77299794-77299894, chr14:77300291-77300391chr14:77300889-77300989, chr14:77301389-77301489, chr14:77301860-77301960, chr14:77302360-77302460, chr14:77302861-77302961, chr14:77303420-77303520, chr14:77303975-77304075, chr14:77304467-77304567, chr14:77304941-77305041, chr14:77305436-77305536, chr14: 77305996-77306096, chr14:77306478-77306578, chr14:77306991-77307091, chr14:77307532-77307632, chr14:77308149-77308249, chr14:77308724-77308824, chr14:77309224-77309324, chr14:77309987-77310087, chr14:77310487-77310587, chr14:77311218-77311318, chr14:773117 18-77311818, chr14:77312218-77312318, chr14:77312718-77312818, chr14:77313138-77313238, chr14:77313591-77313691, chr14:77314092-77314192, chr14:77314616-77314716, chr14:77315105-77315205, chr14:77315604-77315704, chr14:77316187-77316287, chr14:77316687-773 16787, chr14:77317186-77317286, chr14:77317586-77317686, chr14:77318176-77318276, chr14:77318627-77318727, chr14:77319129-77319229, chr14:77319630-77319730, chr14:77320130-77320230, chr14:77320630-77320730, chr14:77321119-77321219, chr14:77321519-77321619chr14:77322079-77322179,chr14:77322579-77322679,chr17:5016400 4-50164104,chr17:50164504-50164604,chr17:50164995-50165095,chr 17:50165501-50165601,chr17:50166047-50166147,chr17:50166483-5 0166583,chr17:50167061-50167161,chr17:50167561-50167661,chr17: 50168002-50168102,chr17:50168502-50168602,chr17:50169002-5016 9102,chr17:50169514-50169614,chr17:50169980-50170080,chr17:501 70511-50170611,chr17:50171060-50171160,chr17:50171575-5017167 5,chr17:50172083-50172183,chr17:50172627-50172727,chr17:501730 70-50173170,chr17:50173570-50173670,chr17:50174370-50174470,c hr17:50174770-50174870,chr17:50175270-50175370,chr17:50175706- 50175806, chr17:50176280-50176380, chr17:50176823-50176923, chr17:50177248-50177348, chr17:50177745-50177845, chr19:46744076-467 44176, chr19:46744665-46744765, chr19:46745165-46745265, chr19:46745654-46745754, chr19:46746154-46746254, chr19:46746654-467467 54, chr19:46747154-46747254, chr19:46747688-46747788, chr19:46748088-46748188, chr19:46748592-46748692, chr19:46749078-46749178chr19:46750078-46750178, chr19:46750878-46750978, chr19:46751852-46751952, chr19:46752254-46752354, chr19:46753524-46753624, chr 19:46753964-46754064, chr19:46754510-46754610, chr19:46755091-46755191, chr19:46755591-46755691, chr19:46756053-46756153, chr19: 46756584-46756684, chr19:46757056-46757156, chr19:46757556-46757656, chr19:46758051-46758151, chr19:46758489-46758589, chr19:467 59155-46759255, chr19:46759755-46759855, chr19:46760277-46760377, chr3:49467180-49467280, chr3:49467580-49467680, chr3:49468180- 49468280,chr3:49468954-49469054,chr3:49469486-49469586,chr3:4 9469985-49470085,chr3:49470485-49470585,chr3:49470990-49471090 ,chr3:49471445-49471545,chr3:49471925-49472025,chr3:49472425- 49472525,chr3:49473425-49473525,chr3:49475581-49475681,chr3:49 476081-49476181,chr3:49476540-49476640,chr3:49477540-49477640 ,chr3:49478592-49478692,chr3:49479181-49479281,chr3:49480577-4 9480677,chr3:49481501-49481601,chr3:49481929-49482029,chr3:49 482729-49482829,chr3:49483488-49483588,chr3:49483973-49484073,chr3:49484456-49484556,chr3:49485341-49485441,chr3:49485941-49486041,chr3:49486784-49486884,chr3:49487259-49487359,chr3:49488025-49488125,chr3:49488517-49488617,chr3:49488990-49489090,chr3:49489473-49489573,chr3:49489943-49490043,chr3:49490436-49490536,chr3:49492206-49492306,chr3:49492742-49492842,chr3:49493340-49493440,chr3:49493830-49493930,chr3:49494355-49494455,chr3:49494908-49495008,chr3:49495408-49495508,chr3:49495952-49496052,chr3:49496952-49497052,chr3:49497552-49497652,chr3:49498052-49498152,chr3:49498522-49498622,chr3:49499106-49499206,chr3:49499563-49499663,chr3:49500304-49500404,chr3:49500804-49500904,chr3:49501304-49501404,chr3:49501813-49501913,chr3:49502313-49502413,chr3:49502913-49503013,chr3:49503326-49503426,chr3:49503913-49504013,chr3:49504407-49504507,chr3:49505405-49505505,chr3:49506342-49506442,chr3:49506754-49506854,chr3:49507195-49507295,chr3:49507786-49507886,chr3:49508908-49509008,chr3:49509393-49509493,chr3:49509959-49510059,chr3:49510453-49510553,chr3:49510953-49511053,chr3:49511445-49511545,chr3:49511845-49511945,chr3:49512947-49513047,chr3:49513545-49513645,chr3:49514060-49514160,chr3:49515278-49515378,chr3:49515797-49515897,chr3:49516297-49516397,chr3:49516797-49516897,chr3:49517297-49517397,chr3:49517810-49517910,chr3:49518310-49518410,chr3:49518839-49518939,chr3:49519339-49519439,chr3:49519794-49519894,chr3:49520258-49520358,chr3:49520758-49520858,chr3:49521645-49521745,chr3:49522594-49522694,chr3:49523068-49523168,chr3:49523520-49523620,chr3:49524020-49524120,chr3:49524476-49524576,chr3:49525076-49525176,chr3:49526180-49526280,chr3:49526780-49526880,chr3:49527778-49527878,chr3:49528179-49528279,chr3:49528701-49528801,chr3:49529301-49529401,chr3:49529901-49530001,chr3:49530401-49530501,chr3:49530901-49531001,chr3:49531401-49531501,chr3:49531902-49532002,chr3:49532402-49532502,chr3:49532849-49532949,chr3:49533282-49533382,chr3:49533782-49533882,chr3:49534326-49534426,chr3:49534801-49534901,chr3:49535277-49535377,chr3:49535777-49535877,chr3:49536277-49536377,chr3:49536857-49536957,chr3:49537594-49537694,chr3:49717919-49718019,chr3:49718419-49718519,chr3:49718920-49719020,chr3:49719426-49719526,chr3:49719876-49719976,chr3:49720377-49720477,chr3:49720878-49720978,chr3:49721378-49721478,chr3:49721841-49721941,chr3:49722311-49722411,chr3:49722812-49722912,chr3:49723312-49723412,chr3:49723830-49723930,chr3:49724330-49724430,chr3:49724856-49724956,chr3:49725328-49725428,chr3:49725801-49725901,chr4:52018705-52018805,chr4:52019255-52019355,chr4:52019777-52019877,chr4:52020277-52020377,chr4:52020719-52020819,chr4:52021219-52021319,chr4:52021760-52021860,chr4:52022328-52022428,chr4:52022828-52022928,chr4:52023291-52023391,chr4:52023809-52023909,chr4:52024365-52024465,chr4:52024852-52024952,chr4:52025299-52025399,chr4:52025709-52025809,chr4:52026114-52026214,chr4:52026614-52026714,chr4:52027181-52027281,chr4:52027683-52027783,chr4:52028218-52028318,chr4:52028718-52028818,chr4:52029188-52029288,chr4:52029736-52029836,chr4:52030336-52030436,chr4:52030834-52030934,chr4:52031239-52031339,chr4:52031739-52031839,chr4:52 032243-52032343,chr4:52032782-52032882,chr4:52033299-52033399 ,chr4:52033722-52033822,chr4:52034628-52034728,chr4:52035128-5 2035228,chr4:52035622-52035722,chr4:52036124-52036224,chr4:52 036625-52036725,chr4:52037079-52037179,chr4:52037564-52037664, chr4:52037989-52038089,chr4:52038489-52038589,chr4:52039056-52039156,chr4:52039556-52039656,chr4:52040066-52040166,chr5:156 325162-156325262,chr5:156325662-156325762,chr5:156326162-1563 26262,chr5:156326684-156326784,chr5:156327184-156327284,chr5:1 56327663-156327763,chr5:156328163-156328263,chr5:156328670-15 6328770,chr5:156329084-156329184,chr5:156329544-156329644,chr5 :156330031-156330131,chr5:156330570-156330670,chr5:156331054- 156331154,chr5:156331542-156331642,chr5:156332042-156332142,ch r5:156332578-156332678,chr5:156333102-156333202,chr5:15633355 5-156333655,chr5:156334028-156334128,chr5:156334517-156334617,chr5:156335110-156335210, chr5:156335610-156335710, chr5:156336099-156336199, chr5:156336585-156336685, chr5:156337159-156337259 chr5:156337622-156337722, chr5:156338170-156338270, chr5:156338673-156338773, chr5:156339155-156339255, chr5:156339584-156339684 chr5:156340016-156340116, chr5:156340516-156340616, chr5:156341072-156341172, chr5:156341529-156341629, chr5:156342053-156342153 chr5:156342564-156342664, chr5:156343113-156343213, chr5:156343613-156343713, chr5:156344113-156344213, chr5:156344613-156344713 chr5:156345113-156345213, chr5:156345691-156345791, chr5:156346122-156346222, chr5:156346624-156346724, chr5:156347159-156347259 chr5:156347668-156347768, chr5:156348129-156348229, chr5:156348622-156348722, chr5:156349102-156349202, chr5:156349702-156349802 chr5:156350287-156350387, chr5:156350783-156350883, chr5:156351264-156351364, chr5:156351856-156351956, chr5:156352320-156352420 chr5:156352820-156352920, chr5:156353333-156353433, chr5:156353833-156353933, chr5:156354370-156354470, chr5:156354970-156355070chr5:156355493-156355593, chr5:156356002-156356102, chr5:156356541-156356641, chr5:156357061-156357161, chr5:156357467-156357567 chr5:156357974-156358074, chr5:156358522-156358622, chr5:156359028-156359128, chr5:156359535-156359635, chr5:156359948-156360048 chr5:156360512-156360612, chr5:156361085-156361185, chr5:156361581-156361681, chr5:156362071-156362171, chr5:156362659-156362759 chr5:156363159-156363259, chr5:156363688-156363788, chr5:156364234-156364334, chr5:156364734-156364834, chr5:156365234-156365334 chr5:156365792-156365892, chr5:156366317-156366417, chr5:156366898-156366998, chr5:156367459-156367559, chr5:156368000-156368100 chr5:156368500-156368600, chr5:156369065-156369165, chr5:156369511-156369611, chr5:156370011-156370111, chr5:156370539-156370639 chr5:156371012-156371112, chr5:156371430-156371530, chr5:156371930-156372030, chr5:156372430-156372530, chr5:156372944-156373044 chr5:156373494-156373594, chr5:156374037-156374137, chr5:156374459-156374559, chr5:156375028-156375128, chr5:156375521-156375621chr5:156376021-156376121, chr5:156376513-156376613, chr5:156377113-156377213, chr5:156377616-156377716, chr5:156378214-156378314 chr5:156378739-156378839, chr5:156379239-156379339, chr5:156379739-156379839, chr5:156380239-156380339, chr5:156380774-156380874 chr5:156381321-156381421, chr5:156381744-156381844, chr5:156382244-156382344, chr5:156382736-156382836, chr5:156383139-156383239 chr5:156383644-156383744, chr5:156384144-156384244, chr5:156384743-156384843, chr5:156385310-156385410, chr5:156385744-156385844 chr5:156386244-156386344, chr5:156386774-156386874, chr5:156387283-156387383, chr5:156387783-156387883, chr5:156388269-156388369 chr5:156388850-156388950, chr5:156389350-156389450, chr5:156389818-156389918, chr5:156390238-156390338, chr5:156390801-156390901 chr5:156391206-156391306, chr5:156391734-156391834, chr5:156392234-156392334, chr5:156392696-156392796, chr5:156393193-156393293 chr5:156393691-156393791, chr5:156394194-156394294, 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chr5:156663775-156663875, chr5:156664341-156664441, chr5:156664941-156665041, chr5:156665364-156665464 chr5:156665764-156665864, chr5:156666964-156667064, chr5:156667564-156667664, chr5:156667964-156668064, chr5:156668384-156668484 chr5:156668884-156668984, chr5:156669299-156669399, chr5:156669881-156669981, chr5:156670370-156670470, chr5:156670872-156670972 chr5:156671596-156671696, chr5:156672195-156672295, chr5:156672756-156672856, chr5:156673332-156673432, chr5:156673782-156673882 chr5:156674253-156674353, chr5:156674753-156674853, chr5:156675352-156675452, chr5:156675908-156676008, chr5:156676415-156676515 chr5:156676915-156677015, chr5:156677715-156677815, chr5:156678195-156678295, chr5:156678695-156678795, chr5:156679231-156679331chr5:156679774-156679874, chr5:156680274-156680374, chr5:156680767-156680867, chr5:156681273-156681373, chr5:156681811-156681911 chr5:156682311-156682411, chr5:156682859-156682959, chr5:156683277-156683377, chr5:156683776-156683876, chr5:156684276-156684376 chr5:156684776-156684876, chr5:156685326-156685426, chr5:156685747-156685847, chr5:156686247-156686347, chr5:156686724-156686824 chr5:156687199-156687299, chr5:156687699-156687799, chr5:156688299-156688399, chr5:156688700-156688800, chr5:156689300-156689400 chr5:156689767-156689867, chr5:156690319-156690419, chr5:156690837-156690937, chr5:156691330-156691430, chr5:156691746-156691846 chr5:156692230-156692330, chr5:156692730-156692830, chr5:156693164-156693264, chr5:156693692-156693792, chr5:156694195-156694295 chr5:156694695-156694795, chr5:156695183-156695283, chr5:156695718-156695818, chr5:156696219-156696319, chr5:156696790-156696890 chr5:156697290-156697390, chr5:156697798-156697898, chr5:156698268-156698368, chr5:156698738-156698838, chr5:156699259-156699359chr5:156699760-156699860, chr5:156700221-156700321, chr5:156700797-156700897, chr5:156701325-156701425, chr5:156701886-156701986 chr5:156702386-156702486, chr5:156702918-156703018, chr5:156703418-156703518, chr5:156703890-156703990, chr5:156704370-156704470 chr5:156704954-156705054, chr5:156705454-156705554, chr5:156705954-156706054, chr5:156706477-156706577, chr5:156706907-156707007 chr5:156707407-156707507, chr5:156707917-156708017, chr5:156708442-156708542, chr5:156708942-156709042, chr5:156709395-156709495 chr5:156709920-156710020, chr5:156710323-156710423, chr5:156710886-156710986, chr5:156711384-156711484, chr5:156712179-156712279 chr5:156712633-156712733, chr5:156713098-156713198, chr5:156713653-156713753, chr5:156714114-156714214, chr5:156714604-156714704 chr5:156715034-156715134, chr5:156715533-156715633, chr5:156716058-156716158, chr5:156716584-156716684, chr5:156717084-156717184 chr5:156717584-156717684, chr5:156718094-156718194, chr5:156718599-156718699, chr5:156719122-156719222, chr5:156719622-156719722chr5:156720102-156720202, chr5:156720601-156720701, chr5:156721064-156721164, chr5:156721561-156721661, chr5:156722061-156722161 chr5:156722558-156722658, chr5:156723046-156723146, chr5:156723537-156723637, chr5:156724037-156724137, chr5:156724616-156724716 chr5:156725149-156725249, chr5:156725641-156725741, chr5:156726146-156726246, chr5:156726652-156726752, chr5:156727107-156727207 chr5:156727607-156727707, chr5:156728148-156728248, chr5:156728654-156728754, chr5:156729154-156729254, chr5:156729705-156729805 chr5:156730145-156730245, chr5:156730759-156730859, chr5:156731899-156731999, chr5:156732602-156732702, chr5:156733119-156733219 chr5:156733900-156734000, chr5:156734449-156734549, chr5:156734861-156734961, chr5:156735445-156735545, chr5:156736001-156736101 chr5:156736528-156736628, chr5:156737004-156737104, chr5:156737507-156737607, chr5:156738035-156738135, chr5:156738507-156738607 chr5:156739004-156739104, chr5:156739504-156739604, chr5:156739972-156740072, chr5:156740472-156740572, chr5:156740972-156741072chr5:156741472-156741572, chr5:156742213-156742313, chr5:156742799-156742899, chr5:156743399-156743499, chr5:156743907-156744007 chr5:156744343-156744443, chr5:156744941-156745041, chr5:156745450-156745550, chr5:156745896-156745996, chr5:156746365-156746465 chr5:156746954-156747054, chr5:156747458-156747558, chr5:156747960-156748060, chr5:156748483-156748583, chr5:156749043-156749143 chr5:156749499-156749599, chr5:156749977-156750077, chr5:156750677-156750777, chr5:156751157-156751257, chr5:156751739-156751839 chr5:156752255-156752355, chr5:156752801-156752901, chr5:156753301-156753401, chr5:156753885-156753985, chr5:156754406-156754506 chr5:156754917-156755017, chr5:156755366-156755466, chr5:156755832-156755932, chr5:156756554-156756654, chr5:156757061-156757161 chr5:156757566-156757666, chr5:156758066-156758166, chr5:156758596-156758696, chr5:156759002-156759102, chr5:156759402-156759502 chr5:156759913-156760013, chr5:156760382-156760482, chr5:156760882-156760982, chr5:156761322-156761422, chr5:156761821-156761921chr5:156762321-156762421, chr5:156762824-156762924, chr5:156763324-156763424, chr5:156763824-156763924, chr5:156764306-156764406 , chr5:156764735-156764835, chr5:156765226-156765326, chr5:156765739-156765839, chr5:156766207-156766307, chr5:156766719-156766819 , chr5:156767236-156767336, chr5:156767755-156767855, chr5:156768255-156768355, chr5:156768747-156768847, chr5:156769293-15676939 3. Chr7:16085530-16085630, Chr7:16086265-16086365, Chr7:16086813-16086913, Chr7:16087373-16087473, Chr7:16087887-16087987, Chr7:160 88317-16088417, chr7:16088817-16088917, chr7:16089267-16089367, chr7:16089818-16089918, chr7:16090306-16090406, chr7:16090853-160 90953, chr7:16091347-16091447, chr7:16091847-16091947, chr7:16092359-16092459, chr7:16092796-16092896, chr7:16093290-16093390, chr7 :16093778-16093878, chr7:16094278-16094378, chr7:16094726-16094826, chr7:16095203-16095303, chr7:16095703-16095803, chr7:16096141 -16096241, chr7:16096697-16096797, chr7:16097197-16097297, chr7:16097673-16097773, chr7:16098173-16098273, chr7:16098738-16098838Chr7:16099295-16099395, Chr7:16099795-16099895, Chr7:16100330-16100430, Chr7:16101545-16101645, Chr7:16103545-16103645, Chr7:161 04067-16104167, chr7:16104501-16104601, chr7:16105016-16105116, chr7:16105502-16105602, chr7:16106002-16106102, chr7:16106420-161 06520, chr7:16107019-16107119, chr7:16107581-16107681, chr7:16108167-16108267, chr7:16108664-16108764, chr7:16109264-16109364, ch r7:16109810-16109910, chr7:16110337-16110437, chr7:16110833-16110933, chr7:16111272-16111372, chr7:16111729-16111829, chr7:161123 26-16112426, chr7:16112756-16112856, chr7:16113203-16113303, chr7:16113647-16113747, chr7:16114167-16114267, chr7:16114700-16114 800, chr7:16115181-16115281, chr7:16115669-16115769, chr7:16116224-16116324, chr7:16116794-16116894, chr7:16117294-16117394, chr7: 16117758-16117858, chr7:16118258-16118358, chr7:16118841-16118941, chr7:16119275-16119375, chr7:16119775-16119875, chr7:16120358- 16120458, chr7:16120880-16120980, chr7:16121307-16121407, chr7:16121813-16121913, chr7:16122313-16122413, chr7:16122911-16123011Chr7:16123396-16123496, Chr7:16123858-16123958, Chr7:16124276-16124376, Chr7:16124876-16124976, Chr7:16125329-16125429, Chr7:161 25799-16125899, chr7:16126299-16126399, chr7:16126771-16126871, chr7:16127263-16127363, chr7:16127862-16127962, chr7:16128400-161 28500, chr7:16128935-16129035, chr7:16129435-16129535, chr7:16129919-16130019, chr7:16130514-16130614, chr7:16130993-16131093, ch r7:16131470-16131570, chr7:16131934-16132034, chr7:16132389-16132489, chr7:16132789-16132889, chr7:16133328-16133428, chr7:161337 84-16133884, chr7:16134286-16134386, chr7:16134788-16134888, chr7:16135293-16135393, chr7:16135785-16135885, chr7:16136373-16136 473, chr7:16136861-16136961, chr7:16137352-16137452, chr7:16137806-16137906, chr7:16138306-16138406, chr7:16138707-16138807, chr7: 16139271-16139371, chr7:16139832-16139932, chr7:16140432-16140532, chr7:16140959-16141059, chr7:16141441-16141541, chr7:16141978- 16142078, chr7:16142478-16142578, chr7:16142939-16143039, chr7:16143446-16143546, chr7:16143897-16143997, chr7:16144300-16144400Chr7:16144763-16144863, Chr7:16145263-16145363, Chr7:16145783-16145883, Chr7:16146231-16146331, Chr7:16146770-16146870, Chr7:161 47270-16147370, chr7:16147744-16147844, chr7:16148151-16148251, chr7:16148683-16148783, chr7:16149238-16149338, chr7:16150009-161 50109, chr7:16150421-16150521, chr7:16150828-16150928, chr7:16151326-16151426, chr7:16151826-16151926, chr7:16152367-16152467, ch r7:16152889-16152989, chr7:16153389-16153489, chr7:16153900-16154000, chr7:16154413-16154513, chr7:16155174-16155274, chr7:161557 68-16155868, chr7:16156268-16156368, chr7:16156792-16156892, chr7:16157330-16157430, chr7:16157747-16157847, chr7:16158280-16158 380, chr7:16158780-16158880, chr7:16159297-16159397, chr7:16161627-16161727, chr7:16163977-16164077, chr7:16164625-16164725, chr7: 16165098-16165198, chr7:16165547-16165647, chr7:16166010-16166110, chr7:16166566-16166666, chr7:16167266-16167366, chr7:16167776- 16167876, chr7:16168301-16168401, chr7:16168820-16168920, chr7:16169335-16169435, chr7:16169907-16170007, chr7:16170607-16170707Chr7:16171177-16171277, Chr7:16171877-16171977, Chr7:16172438-16172538, Chr7:16172919-16173019, Chr7:16173364-16173464, Chr7:161 73768-16173868, chr7:16174199-16174299, chr7:16174716-16174816, chr7:16175170-16175270, chr7:16175633-16175733, chr7:16176038-161 76138, chr7:16176488-16176588, chr7:16177087-16177187, chr7:16177492-16177592, chr7:16177967-16178067, chr7:16178467-16178567, ch r7:16178953-16179053, chr7:16179430-16179530, chr7:16179974-16180074, chr7:16180474-16180574, chr7:16180948-16181048, chr7:161814 78-16181578, chr7:16182011-16182111, chr7:16182525-16182625, chr7:16183052-16183152, chr7:16183552-16183652, chr7:16184304-16184 404, chr7:16184814-16184914, chr7:16185414-16185514, chr7:16185914-16186014, chr7:16186319-16186419, chr7:16186819-16186919, chr7: 16187401-16187501, chr7:16187814-16187914, chr7:16188292-16188392, chr7:16188837-16188937, chr7:16189386-16189486, chr7:16189886- 16189986, chr7:16190286-16190386, chr7:16190842-16190942, chr7:16191353-16191453, chr7:16191873-16191973, chr7:16192345-16192445chr7:16192902-16193002, chr7:16193441-16193541, chr7:16193941-16194041, chr7:16194441-16194541, chr7:16194939-16195039, chr7:161 95401-16195501, chr7:16195822-16195922, chr7:16196308-16196408, chr7:16196793-16196893, chr7:16197197-16197297, chr7:16197637-161 97737, chr7:16198817-16198917, chr7:16199217-16199317, chr7:16199747-16199847, chr7:16200317-16200417, chr7:16200846-16200946, ch r7:16201321-16201421, chr7:16201909-16202009, chr7:16202412-16202512, chr7:16202974-16203074, chr7:16203474-16203574, chr7:162040 25-16204125, chr7:16204625-16204725, chr7:16205126-16205226, chr7:16205640-16205740, chr7:16206192-16206292, chr7:16206785-16206 885, chr7:16207274-16207374, chr7:16207839-16207939, chr7:16208286-16208386, chr7:16208751-16208851, chr7:16209181-16209281, chr7: 16209645-16209745, chr7:16210162-16210262, chr7:16210691-16210791, chr7:16211147-16211247, chr7:16211634-16211734, chr7:16212134- 16212234, chr7:16212636-16212736, chr7:16213136-16213236, chr7:16213836-16213936, chr7:16214288-16214388, chr7:16214788-16214888chr7:16215283-16215383, chr7:16215801-16215901, chr7:16216305-16216405, chr7:16222480-16222580, chr7:16222921-16223021, chr7:162 23421-16223521, chr7:16223885-16223985, chr7:16224385-16224485, chr7:16224905-16225005, chr7:16225405-16225505, chr7:16225967-162 26067, chr7:16226467-16226567, chr7:16226874-16226974, chr7:16227280-16227380, chr7:16227813-16227913, chr7:16228216-16228316, ch r7:16228802-16228902, chr7:16229322-16229422, chr7:16229749-16229849, chr7:16230216-16230316, chr7:16230739-16230839, chr7:162311 39-16231239, chr7:16231700-16231800, chr7:16232138-16232238, chr7:16232547-16232647, chr7:16233147-16233247, chr7:16233614-16233 714, chr7:16234149-16234249, chr7:16234641-16234741, chr7:16235132-16235232, chr7:16235632-16235732, chr7:16236203-16236303, chr7: 16236625-16236725, chr7:16237141-16237241, chr7:16237699-16237799, chr7:16238226-16238326, chr7:16238741-16238841, chr7:16239231- 16239331, chr7:16239670-16239770, chr7:16240161-16240261, chr7:16240697-16240797, chr7:16241245-16241345, chr7:16241809-16241909chr7:16242343-16242443, chr7:16242812-16242912, chr7:16243359-16243459, chr7:16243959-16244059, chr7:16244488-16244588, chr7:162 44934-16245034, chr7:16245434-16245534, chr7:16245901-16246001, chr7:16246346-16246446, chr7:16246870-16246970, chr7:16247463-162 47563, chr7:16247963-16248063, chr7:16248478-16248578, chr7:16249060-16249160, chr7:16249551-16249651, chr7:16250073-16250173, ch r7:16252089-16252189, chr7:16252808-16252908, chr7:16256973-16257073, chr7:16257482-16257582, chr7:16257982-16258082, chr7:162584 92-16258592, chr7:16258978-16259078, chr7:16259489-16259589, chr7:16260080-16260180, chr7:16260564-16260664, chr7:16261064-16261 164, chr7:16261640-16261740, chr7:16262085-16262185, chr7:16262560-16262660, chr7:16263159-16263259, chr7:16263759-16263859, chr7: 16264232-16264332, chr7:16264730-16264830, chr7:16265241-16265341, chr7:16265741-16265841, chr7:16266241-16266341, chr7:16266818- 16266918, chr7:16267317-16267417, chr7:16267728-16267828, chr7:16268278-16268378, chr7:16268778-16268878, chr7:16269303-16269403chr7:16269806-16269906, chr7:16270207-16270307, ​​chr7:16270800-16270900, chr7:16271299-16271399, chr7:16271797-16271897, chr7:162 72316-16272416, chr7:16272780-16272880, chr7:16273285-16273385, chr7:16273788-16273888, chr7:16274333-16274433, chr7:16274804-162 74904, chr7:16275305-16275405, chr7:16275870-16275970, chr7:16276349-16276449, chr7:16276898-16276998, chr7:16277398-16277498, ch r7:16277890-16277990, chr7:16278381-16278481, chr7:16278842-16278942, chr7:16279342-16279442, chr7:16279744-16279844, chr7:162802 34-16280334, chr7:16280643-16280743, chr7:16281055-16281155, chr7:16281548-16281648, chr7:16282148-16282248, chr7:16282659-16282 759, chr7:16283093-16283193, chr7:16283611-16283711, chr7:16284111-16284211, chr7:16284611-16284711, chr7:16285143-16285243, chr7: 16285643-16285743, chr7:16286243-16286343, chr7:16286736-16286836, chr7:16287289-16287389, chr7:16287956-16288056, chr7:16288396- 16288496, chr7:16288908-16289008, chr7:16289381-16289481, chr7:16289942-16290042, chr7:16290521-16290621, chr7:16291118-16291218chr7:16291521-16291621, chr7:16292021-16292121, chr7:16292594-16292694, chr7:16293144-16293244, chr7:16293644-16293744, chr7:162 94139-16294239, chr7:16294639-16294739, chr7:16295139-16295239, chr7:16295657-16295757, chr7:16296171-16296271, chr7:16296600-162 96700, chr7:16297100-16297200, chr7:16297628-16297728, chr7:16298204-16298304, chr7:16298712-16298812, chr7:16299212-16299312, ch r7:16299734-16299834, chr7:16300289-16300389, chr7:16300872-16300972, chr7:16301382-16301482, chr7:16301871-16301971, chr7:163023 27-16302427, chr7:16302833-16302933, chr7:16303233-16303333, chr7:16303680-16303780, chr7:16304223-16304323, chr7:16304792-16304 892, chr7:16305358-16305458, chr7:16305907-16306007, chr7:16306338-16306438, chr7:16306782-16306882, chr7:16307276-16307376, chr7: 16307762-16307862, chr7:16308319-16308419, chr7:16308786-16308886, chr7:16309286-16309386, chr7:16309830-16309930, chr7:16310330- 16310430, chr7:16310766-16310866, chr7:16311266-16311366, chr7:16311860-16311960, chr7:16312386-16312486, chr7:16312886-16312986Chr7:16313286-16313386, Chr7:16313686-16313786, Chr7:16314283-16314383, Chr7:16314722-16314822, Chr7:16315313-16315413, Chr7:163 15764-16315864, chr7:16316248-16316348, chr7:16316842-16316942, chr7:16317329-16317429, chr7:16317927-16318027, chr7:16318426-163 18526, chr7:16318926-16319026, chr7:16319426-16319526, chr7:16319939-16320039, chr7:16320439-16320539, chr7:16320939-16321039, ch r7:16321452-16321552, chr7:16321945-16322045, chr7:16322474-16322574, chr7:16323010-16323110, chr7:16323519-16323619, chr7:163240 19-16324119, chr7:16324536-16324636, chr7:16325120-16325220, chr7:16325542-16325642, chr7:16326066-16326166, chr7:16326610-16326 710, chr7:16327159-16327259, chr7:16327620-16327720, chr7:16328080-16328180, chr7:16328859-16328959, chr7:16329449-16329549, chr7: 16329949-16330049, chr7:16330478-16330578, chr7:16330882-16330982, chr7:16331364-16331464, chr7:16331945-16332045, chr7:16332544- 16332644, chr7:16333053-16333153, chr7:16333508-16333608, chr7:16334040-16334140, chr7:16334540-16334640, chr7:16334952-16335052Chr7:16335511-16335611, Chr7:16336107-16336207, Chr7:16336607-16336707, Chr7:16337168-16337268, Chr7:16337601-16337701, Chr7:163 38078-16338178, chr7:16338540-16338640, chr7:16339088-16339188, chr7:16339545-16339645, chr7:16340142-16340242, chr7:16340733-163 40833, chr7:16341309-16341409, chr7:16341770-16341870, chr7:16342229-16342329, chr7:16342829-16342929, chr7:16343345-16343445, ch r7:16343905-16344005, chr7:16344502-16344602, chr7:16344902-16345002, chr7:16345339-16345439, chr7:16345790-16345890, chr7:163463 10-16346410, chr7:16346868-16346968, chr7:16347387-16347487, chr7:16347821-16347921, chr7:16348334-16348434, chr7:16348787-16348 887, chr7:16349307-16349407, chr7:16349835-16349935, chr7:16350335-16350435, chr7:16350760-16350860, chr7:16352227-16352327, chr7: 16352739-16352839, chr7:16353322-16353422, chr7:16353865-16353965, chr7:16354351-16354451, chr7:16354856-16354956, chr7:16355342- 16355442, chr7:16355842-16355942, chr7:16356250-16356350, chr7:16356650-16356750, chr7:16357064-16357164, chr7:16357568-16357668chr7:16358121-16358221, chr7:16358650-16358750, chr7:16359050-16359150, chr7:16359544-16359644, chr7:16360044-16360144, chr7:163 60509-16360609, chr7:16360993-16361093, chr7:16361393-16361493, chr7:16361793-16361893, chr7:16362386-16362486, chr7:16362819-163 62919, chr7:16363386-16363486, chr7:16363865-16363965, chr7:16364286-16364386, chr7:16364780-16364880, chr7:16365265-16365365, ch r7:16365731-16365831, chr7:16366264-16366364, chr7:16366807-16366907, chr7:16367254-16367354, chr7:16367754-16367854, chr7:163681 60-16368260, chr7:16368595-16368695, chr7:16368996-16369096, chr7:16369520-16369620, chr7:16370090-16370190, chr7:16370600-16370 700, chr7:16371172-16371272, chr7:16371652-16371752, chr7:16372212-16372312, chr7:16372812-16372912, chr7:16373377-16373477, chr7: 16373897-16373997, chr7:16374308-16374408, chr7:16374821-16374921, chr7:16375300-16375400, chr7:16375737-16375837, chr7:16376251- 16376351, chr7:16376768-16376868, chr7:16377268-16377368, chr7:16377768-16377868, chr7:16384128-16384228, chr7:16384628-16384728chr7:16385119-16385219, chr7:16385557-16385657, chr7:16386114-16386214, chr7:16386553-16386653, chr7:16387310-16387410, chr7:163 87910-16388010, chr7:16388419-16388519, chr7:16388968-16389068, chr7:16389542-16389642, chr7:16390033-16390133, chr7:16390545-163 90645, chr7:16391015-16391115, chr7:16391515-16391615, chr7:16391963-16392063, chr7:16392452-16392552, chr7:16392995-16393095, ch r7:16393500-16393600, chr7:16394000-16394100, chr7:16394500-16394600, chr7:16395011-16395111, chr7:16395426-16395526, chr7:163959 26-16396026, chr7:16396448-16396548, chr7:16396949-16397049, chr7:16397451-16397551, chr7:16397951-16398051, chr7:16398450-16398 550, chr7:16398951-16399051, chr7:16399451-16399551, chr7:16399952-16400052, chr7:16400483-16400583, chr7:16400983-16401083, chr7: 16401483-16401583, chr7:16401939-16402039, chr7:16402494-16402594, chr7:16402992-16403092, chr7:16403457-16403557, chr7:16403925- 16404025, chr7:16404425-16404525, chr7:16404963-16405063, chr7:16405421-16405521, chr7:16405968-16406068, chr7:16406468-16406568Chr7:16406874-16406974, Chr7:16407382-16407482, Chr7:16408182-16408282, Chr7:16408592-16408692, Chr7:16409165-16409265, Chr7:164 09665-16409765, chr7:16410176-16410276, chr7:16410623-16410723, chr7:16411072-16411172, chr7:16411572-16411672, chr7:16412004-164 12104, chr7:16412504-16412604, chr7:16412978-16413078, chr7:16413544-16413644, chr7:16414044-16414144, chr7:16414529-16414629, ch r7:16415029-16415129, chr7:16415483-16415583, chr7:16415990-16416090, chr7:16416390-16416490, chr7:16416828-16416928, chr7:164172 97-16417397, chr7:16417897-16417997, chr7:16418302-16418402, chr7:16418883-16418983, chr7:16419284-16419384, chr7:16419805-16419 905, chr7:16420305-16420405, chr7:16420805-16420905, chr7:16421305-16421405, chr7:16421805-16421905, chr7:16422284-16422384, chr7: 16422749-16422849, chr7:16423208-16423308, chr8:43091521-43091621, chr8:43092120-43092220, chr8:43092625-43092725, chr8:43093125- 43093225,chr8:43093625-43093725,chr8:43094391-43094491,chr8:43 094923-43095023,chr8:43095512-43095612,chr8:43096007-43096107,chr8:43096707-43096807,chr8:43097297-43097397,chr8:43097823-43097923,chr8:43098223-43098323,chr8:43098788-43098888,chr8:43099310-43099410,chr8:43099795-43099895,chr8:43100317-43100417,chr8:43100817-43100917,chr8:43101542-43101642,chr8:43102139-43102239,chr8:43102639-43102739,chr8:43103206-43103306,chr8:43103745-43103845,chr8:43104370-43104470,chr8:43104957-43105057,chr8:43105357-43105457,chr8:43106312-43106412,chr8:43106880-43106980,chr8:43107290-43107390,chr8:43108059-43108159,chr8:43108521-43108621,chr8:43108999-43109099,chr8:43109427-43109527,chr8:43110007-43110107,chr8:43110465-43110565,chr8:43114815-43114915,chr8:43115311-43115411,chr8:43115775-43115875,chr8:43116186-43116286,chr8:43116673-43116773,chr8:43117273-43117373,chr8:43117822-43117922,chr8:43118290-43118390,chr8:43118714-43118814,chr8:43119214-43119314,chr8:43119741-43119841,chr8:43120507-43120607,chr8:43121019-43121119,chr8:43121561-43121661,chr8:43122076-43122176,chr8:43122597-43122697,chr8:43122997-43123097,Chr8:43123595-43123695, Chr8:43124022-43124122, Chr8:43124482-43124582, Chr8:43124903-43125003, Chr9:105556158-105556258, Chr9:10 5556694-105556794, chr9:105557149-105557249, chr9:105557649-105557749, chr9:105558102-105558202, chr9:105558640-105558740, chr9:1 05559140-105559240, chr9:105559709-105559809, chr9:105560170-105560270, chr9:105560682-105560782, chr9:105561270-105561370, chr9: 105561678-105561778, chr9:105562139-105562239, chr9:105562693-105562793, chr9:105563093-105563193, chr9:105565154-105565254, chr9 :105569354-105569454, chr9:105569872-105569972, chr9:105570360-105570460, chr9:105570915-105571015, chr9:105571339-105571439, chr 9:105571894-105571994, chr9:105572362-105572462, chr9:105572862-105572962, chr9:105573264-105573364, chr9:105573759-105573859, ch r9:105574299-105574399, chr9:105574773-105574873, chr9:105575272-105575372, chr9:105575863-105575963, chr9:105582462-105582562, c hr9:105582962-105583062, chr9:105583475-105583575, chr9:105583981-105584081, chr9:105584413-105584513, chr9:105585013-105585113chr9:105585533-105585633, chr9:105586033-105586133, chr9:105586533-105586633, chr9:105587034-105587134, chr9:105587527-105587627 chr9:105588045-105588145, chr9:105588613-105588713, chr9:105589015-105589115, chr9:105589515-105589615, chr9:105590009-105590109 chr9:105590522-105590622, chr9:105591445-105591545, chr9:105591977-105592077, chr9:105592707-105592807, chr9:105593186-105593286 chr9:105593733-105593833, chr9:105594183-105594283, chr9:105594765-105594865, chr9:105595356-105595456, chr9:105595833-105595933 chr9:105596381-105596481, chr9:105596781-105596881, chr9:105597251-105597351, chr9:105597760-105597860, chr9:105598304-105598404 chr9:105598784-105598884, chr9:105599284-105599384, chr9:105599773-105599873, chr9:105600320-105600420, chr9:105600819-105600919 chr9:105601274-105601374, chr9:105601719-105601819, chr9:105602225-105602325, chr9:105602808-105602908, chr9:105603377-105603477 chr9:105603937-105604037, chr9:105604423-105604523, chr9:105605021-105605121, chr9:105605430-105605530, chr9:105606029-105606129chr9:105606521-105606621, chr9:105607053-105607153, chr9:105607477-105607577, chr9:105607877-105607977, chr9:105608367-105608467 chr9:105608820-105608920, chr9:105609306-105609406, chr9:105609779-105609879, chr9:105610338-105610438, chr9:105610901-105611001 chr9:105611301-105611401, chr9:105611785-105611885, chr9:105612258-105612358, chr9:105612976-105613076, chr9:105613455-105613555 chr9:105613956-105614056, chr9:105614446-105614546, chr9:105615189-105615289, chr9:105615591-105615691, chr9:105616168-105616268 chr9:105616668-105616768, chr9:105617113-105617213, chr9:105617549-105617649, chr9:105617978-105618078, chr9:105618556-105618656 chr9:105619143-105619243, chr9:105619558-105619658, chr9:105620158-105620258, chr9:105620685-105620785, chr9:105621185-105621285 chr9:105621641-105621741, chr9:105622135-105622235, chr9:105622635-105622735, chr9:105623071-105623171, chr9:105623571-105623671 chr9:105623974-105624074, chr9:105624500-105624600, chr9:105625018-105625118, chr9:105625518-105625618, chr9:105625951-105626051chr9:105626363-105626463, chr9:105626763-105626863, chr9:105627265-105627365, chr9:105627764-105627864, chr9:105628264-105628364 chr9:105628794-105628894, chr9:105629294-105629394, chr9:105630213-105630313, chr9:105630630-105630730, chr9:105631145-105631245 chr9:105632457-105632557, chr9:105632956-105633056, chr9:105633456-105633556, chr9:105633900-105634000, chr9:105634420-105634520 chr9:105634920-105635020, chr9:105635389-105635489, chr9:105635863-105635963, chr9:105636361-105636461, chr9:105636821-105636921 chr9:105637276-105637376, chr9:105637738-105637838, chr9:105638237-105638337, chr9:105638737-105638837, chr9:105639228-105639328 chr9:105639817-105639917, chr9:105640600-105640700, chr9:105641128-105641228, chr9:105641628-105641728, chr9:105642151-105642251 chr9:105642551-105642651, chr9:105643146-105643246, chr9:131500917-131501017, chr9:131501417-131501517, chr9:131502072-131502172 chr9:131502615-131502715, chr9:131503115-131503215, chr9:131503615-131503715, chr9:131504115-131504215, chr9:131504556-131504656chr9:131505118-131505218, chr9:131505618-131505718, chr9:131506150-131506250, chr9:131506650-131506750, chr9:131507186-131507286 chr9:131507657-131507757, chr9:131508748-131508848, chr9:131509437-131509537, chr9:131509939-131510039, chr9:131510426-131510526 chr9:131510910-131511010, chr9:131511418-131511518, chr9:131512000-131512100, chr9:131512478-131512578, chr9:131512978-131513078 chr9:131513478-131513578, chr9:131513921-131514021, chr9:131514421-131514521, chr9:131514955-131515055, chr9:131515436-131515536 chr9:131515849-131515949, chr9:131516400-131516500, chr9:131516899-131516999, chr9:131517354-131517454, chr9:131517778-131517878 chr9:131518278-131518378, chr9:131518778-131518878, chr9:131519278-131519378, chr9:131519778-131519878, chr9:131520278-131520378 chr9:131520709-131520809, chr9:131521306-131521406, chr9:131521706-131521806, chr9:131522198-131522298, chr9:131522680-131522780 chr9:131523180-131523280, chr9:131523668-131523768, chr9:131524168-131524268, chr9:131524668-131524768, chr9:131525188-131525288chr9:131525779-131525879;, (5) Specific probe sequences for the probe set targeting mitochondrial gene regions may include: chrM: 41-141, chrM: 451-551, chrM: 937-1037, chrM: 1360-1460, chrM: 1807-1907, chrM: 2207-2307, chrM: 2807-2907, chrM: 3298-3398, ... rM: 3709-3809, chrM: 4174-4274, chrM: 4689-4789, chrM: 5289-5389, chrM: 5762-586 2, chrM: 6208-6308, chrM: 6608-6708, chrM: 7117-7217, chrM: 7625-7725, chrM: 8122- 8222, chrM: 8637-8737, chrM: 9040-9140, chrM: 9472-9572, chrM: 9896-9996, chrM: 1 0346-10446, chrM: 10895-10995, chrM: 11318-11418, chrM: 11747-11847, chrM: 12326 -12426, chrM: 12823-12923, chrM: 13320-13420, chrM: 13739-13839, chrM: 14251-14 351, chrM: 14743-14843, chrM: 15269-15369, chrM: 15806-15906, chrM: 16246-16346; (6) The specific probe sequences of the probe set targeting the sex identification gene region may include: chrY:2784852-2784952, chrY:2785491-2785591, chrY:2785995-2786095, chrY:2786495-2786595, chrY:2786995-2787095, chrY:2787495-2787595, chrY:2788002-2788102, chrY:2788494-2788594, chrY:2788994-2789094, chrY:2789494-2789594.

[0025] On the other hand, the present invention provides a kit comprising the probe combination described herein.

[0026] In some implementations, the kit further includes pre-capture library construction reagents; hybridization capture library construction reagents; post-capture PCR amplification reagents; and third-generation library construction reagents.

[0027] In some embodiments, the pre-capture library construction reagent includes a repair enzyme, a repair buffer, a ligase, a ligation buffer, and a adapter. In some embodiments, the adapter is a Y-type adapter, a sticky-end adapter, or a blunt-end adapter.

[0028] In some embodiments, the hybridization capture library construction reagent includes hybridization buffer, hybridization enhancement buffer, human cot-1 DNA, capture magnetic beads, capture washing reagent, and optional sequence blocking reagent. In some embodiments, the sequence blocking reagent contains a sequence that is inversely complementary to the sequence of the adapter. In some embodiments, the capture magnetic beads are streptavidin beads. In some embodiments, no sequence blocking reagent is used.

[0029] In some embodiments, the post-capture PCR amplification reagent includes an amplification enzyme, an amplification buffer, a PCR additive, and amplification primers. In some embodiments, the amplification enzyme is suitable for amplifying long fragments and meets high-fidelity requirements, exhibiting good amplification uniformity and enabling the amplification of high-GC / AT template fragments. In some embodiments, the PCR additive can reduce secondary structure, enhance enzyme stability, or inhibit nonspecific amplification, and is selected from at least one of dimethyl sulfoxide, betaine, bovine serum albumin, and formamide.

[0030] In some embodiments, the third-generation library construction reagent includes a repair enzyme, a repair buffer, a ligase, a ligation buffer, an exonuclease, an exonuclease buffer, a third-generation adapter, and a chip select reagent. In some embodiments, the chip select reagent includes magnetic beads and Buffer EB.

[0031] In another aspect, the present invention provides a method for constructing a single-molecule long-read sequencing library, comprising the following steps: S1) extracting DNA from the sample to be tested and fragmenting it; S2) performing end repair and adapter ligation on the fragmented DNA to obtain a pre-capture library; S3) hybridizing the pre-capture library with the probe combination described herein for capture and PCR amplification to obtain a post-capture PCR product; S4) performing third-generation library construction on the post-capture PCR product to obtain a single-molecule long-read sequencing library.

[0032] In some implementations, the sample to be tested includes blood, dried blood spots, amniotic fluid, cultured amniotic fluid cells, hair, saliva, tissue, semen, sperm stains, and bone.

[0033] In some embodiments, the fragmentation includes sonication, mechanical breaking, or enzymatic digestion. In some embodiments, the fragmented DNA is 7-15 kb in length.

[0034] In some implementations, the adapter for the pre-capture library is prepared by annealing two single strands of DNA and can be a Y-type adapter, a sticky adapter, or a blunt-end adapter. The adapter sequence can be AATGATACGGCGACCACCGAGATNNNNNNNNNNTCCCTACACGACGCTCTTCCGATC (SEQ ID NO: 1) or GATCGGAAGAGCACACGTCTGAACNNNNNNNNNNTCTCGTATGCCGTCTTCTGCTTG (SEQ ID NO: 2). The N part is used to identify the different samples constructed.

[0035] In some implementations, step S2 can be performed without PCR amplification to obtain the pre-capture library.

[0036] In some implementations, after end repair and adapter ligation in step S2, PCR amplification is also included to obtain a pre-capture library.

[0037] In some embodiments, the hybridization capture system in step S3 includes hybridization buffer, hybridization enhancement solution, human cot-1 DNA, probe, and optional blocking sequence reagent. In some embodiments, no blocking sequence is added. In some embodiments, a blocking sequence is added, which is reverse complementary to the adapter sequence.

[0038] In some embodiments, the post-capture PCR amplification system in step S3 includes amplification enzyme, amplification buffer, PCR additives, and amplification primers. In some embodiments, samples with high DNA input may be exempt from amplification. In some embodiments, the amplification primers have a final concentration range of 0.1-1 μM, are phosphorylated, and are adapted to pre-library adapters. In some embodiments, the amplification primers consist of an upstream F primer and a downstream R primer, with nucleotide sequences of AATGATACGGCGACCACCGAGAT (SEQ ID NO: 3) and CAAGCAGAAGACGGCATACGAG (SEQ ID NO: 4). The amplification program includes 94 ℃ for 2-4 min; 98 ℃ for 10-15 sec, Tm ℃ for 30 sec, 68 ℃ for 5-10 min, 15-25 cycles; and 68 ℃ for 5-10 min.

[0039] In some implementations, the system for constructing the third-generation sequencing library in step S4 includes a repair enzyme, a repair enzyme buffer, a ligase, a buffer, an exonuclease, an exonuclease buffer, and a suitable third-generation sequencing adapter.

[0040] In some implementations, the method further includes step S5) performing slice selection on the single-molecule long-read sequencing library, wherein the slice selection reagents include magnetic beads and Buffer EB.

[0041] In another aspect, the present invention provides a method for detecting DMD Gene, DMD The device for modifying genes and identifying and diagnosing genes includes: 1) a pre-capture library construction module: used to fragment the DNA of the sample to be tested and obtain a pre-capture library; 2) a hybridization capture module: used to hybridize and capture the pre-capture library with the probes described herein and amplify it; 3) a third-generation sequencing library construction module: used to construct a single-molecule long-read sequencing library; 4) a sequencing and analysis module: used to sequence on a third-generation sequencing platform and analyze variation information.

[0042] In some implementation schemes, the pre-capture library, post-capture amplification product, and third-generation sequencing library are all obtained according to the method for constructing single-molecule long-read sequencing libraries described herein.

[0043] In some implementations, the third-generation sequencing platform is selected from nanopore-based single-molecule sequencing platforms and single-molecule real-time sequencing platforms.

[0044] In some implementations, the analysis of variant information includes the following steps: i) splitting the sequencing data of the sample to be tested and comparing it with the human reference genome; ii) analyzing the relevant variants, including SNVs, Indels, and SVs, based on the comparison data, and annotating the variants to obtain a variant pathogenicity rating.

[0045] In some implementation schemes, the specific methods for detecting variants are as follows: 1) Analyze variants based on breakpoints, and detect and annotate del and dup information. Simultaneously analyze bnd information to detect inversions, translocations, and complex structural rearrangements; 2) Annotate detected SNVs and Indel sites based on alignment with the reference genome, and interpret the pathogenicity rating.

[0046] In some implementations, the human reference genome is human reference genome version GRCh38.

[0047] In some implementations, the device can perform the following through the above process: a) analyzing and annotating SNV variations of exons; b) analyzing and annotating SNV variations of introns for deep intron variation analysis prediction and annotation; c) analyzing and detecting breakpoint locations and detecting scattered duplicate samples of DMD; d) analyzing and detecting breakpoint locations and detecting complex rearrangement samples.

[0048] In another aspect, the present invention provides the application of the probe combination described herein in the preparation of kits for the diagnosis of Duchenne muscular dystrophy, detection of differential gene variants, differential diagnosis of muscular dystrophy, detection of DMD-related modified gene variants, and detection of DMD transcriptional regulatory region variants.

[0049] The probe combination provided by this invention can simultaneously satisfy DMD Mutation detection DMD Core control area detection, DMD For the detection of modified genes and the identification of diagnostic genes, the probe sets can be combined to meet different clinical application scenarios.

[0050] Compared to existing technologies, this invention has the following advantages: DMD The mutation detection process added DMD Modified genes, DMD Detection of the core regulatory region enables the assessment of Duchenne muscular dystrophy progression, providing crucial guidance for subsequent treatment and disease progression. Simultaneously, this invention enables differential diagnostic analysis, offering further diagnostic references for subjects without available diagnostic information, significantly improving diagnostic efficiency and reducing the risk of missed diagnoses. Attached Figure Description

[0051] Figure 1 The specific detection results for peripheral blood sample No. 30 are: chrX:32557994:[chrX:33166824[A:bnd].

[0052] Figure 2 The specific detection results are for peripheral blood sample number 30: chrX:33018642:T]chrX:33307871]:bnd.

[0053] Figure 3 This is the first complex variant inferred from the breakpoint of peripheral blood sample No. 30.

[0054] Figure 4 This is the second complex variant inferred from the breakpoint of peripheral blood sample No. 30.

[0055] Figure 5 This is the third complex variant inferred from the breakpoint of peripheral blood sample No. 30. Detailed Implementation

[0056] The invention generally described herein will be more readily understood by referring to the following embodiments. Those skilled in the art can use this content to appropriately modify the process parameters for implementation. It should be particularly noted that all similar substitutions and modifications will be obvious to those skilled in the art and are considered to be within the scope of protection of this invention. The following embodiments are provided by way of illustration and are not intended to limit the invention. These embodiments are not intended to represent all or only the experiments conducted below.

[0057] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; and the materials and reagents used are commercially available unless otherwise specified.

[0058] Table 1: Gene Locus Information

[0059] Example 1: Construction of three generations of single-molecule long-read libraries and testing the effect of genomic DNA fragmentation conditions on the average library length. 1. Genomic DNA fragmentation The extracted DNA (1 μg) was fragmented using a fragmentation tube at 8000 rpm, 6000 rpm, and 4000 rpm. After fragmentation, the DNA fragments were purified using AMPure PB magnetic beads (Pacbio, catalog number 100-265-900) to obtain the fragmented DNA.

[0060] 2. DNA Repair Fragmented DNA was repaired using a repair enzyme (Enzymatics, catalog number Y9140-HC-L), and the repair system is shown in Table 2 below. The DNA repair reaction program was 25 °C for 30 minutes; 75 °C for 20 minutes; and held at 4 °C.

[0061] Table 2

[0062] 3. Pre-document library connector connection Phosphorylated adapter material (IDT) was annealed and then formulated into pre-library adapters. The ligation reaction was carried out using ligase (Enzymatics, catalog number L6030-HC-L), and the reaction system is shown in Table 3 below. The DNA ligation reaction program was 25 °C for 30 minutes; then held at 4 °C to obtain the pre-capture library.

[0063] Table 3

[0064] 4. Hybridization of the pre-capture library The hybridization reagent (IDT, catalog number 1072281) was mixed with the purified pre-capture library and 2 μL of the probe combination (D) described in this invention (i.e., probe group (1) + probe group (2) + probe group (3) + probe group (6)). The specific reaction system is shown in Table 4 below. The reaction program was 95℃ for 5 minutes and then maintained at 65℃ for 12-17 hours.

[0065] Table 4

[0066] 5. Capture and product cleaning The hybridization library was thoroughly mixed with streptavidin-labeled capture beads (ThermoFishere, catalog number 65602). After capture incubation, the library was washed using the accompanying hybridization washing kit (IDT, catalog number 1072281). The washed capture beads were then resuspended in enzyme-free water.

[0067] 6. PCR amplification and enrichment of the captured products Prepare the PCR amplification system (TOYOBO, catalog number KFX-201) as shown in Table 5. Add the amplification system to the capture product, mix thoroughly, and then perform PCR amplification according to the reaction procedure shown in Table 6. The amplified product was purified using AMPure PB magnetic beads.

[0068] Table 5

[0069] Table 6

[0070] 7. Construction of third-generation sequencing libraries The single-molecule long-read library construction kit (PacBio, catalog number 103-517-600) was used to construct third-generation libraries. The kit includes ligase, third-generation sequencing adapters, repair enzymes, exonucleases, and corresponding buffer solutions for each component.

[0071] 8. QC results generated from document library data The sequences produced by sequencing were split and aligned, and the average length of the sequences was calculated (Table 7). The results showed that different DNA fragmentation conditions had little effect on the average length of the library.

[0072] Table 7

[0073] Example 2: Construction of third-generation sequencing libraries and library slice selection before sequencing Following the steps in Example 1, third-generation libraries A and B were constructed. Before processing the libraries, slide selection was performed using magnetic beads (AMPure® PB and Auto-Mag®) and Buffer EB. The slide selection results are shown in Table 8.

[0074] Table 8

[0075] This invention adds a slice selection step before library sequencing, which is simple to operate, time-saving, and effectively increases the average length of the library. Increased average length facilitates variant detection and yields more data. The increased average length allows for higher sequencing depth with the same amount of samples, improving the detection of variant sites, especially breakpoints in complex samples. Furthermore, the increased average sequence length leads to longer phase blocks during haplotype analysis, further enhancing variant analysis.

[0076] Example 3: Constructing a third-generation sequencing library and validating the performance of variant detection Following the library construction methods in Examples 1 and 2, DNA samples from six national reference cell lines for preimplantation genetic mutation detection in Duchenne muscular dystrophy (D011212, D011213, D011233, D011242, D011243, D102623) and one Coriell human genome standard cell line DNA sample (GM12878) were sequenced and analyzed. The types of mutations and their pathogenicity ratings were analyzed (Table 9). The results showed that the detection information of the six national reference samples was consistent with the description.

[0077] Table 9: Detection of Duchenne Muscular Dystrophy Gene Mutation in Preimplantation Genes (NPC Reference Material)

[0078] Following the library construction methods of Examples 1 and 2, 30 clinical peripheral blood DNA samples were sequenced and analyzed to determine the types and pathogenicity ratings of variants (Table 10). The probe combinations used are shown in the rightmost column of Table 10, where probe combination A is probe group (1) + probe group (6); probe combination B is probe group (1) + probe group (2) + probe group (3) + probe group (4) + probe group (5) + probe group (6). The results showed that 14 of the 30 clinical samples had the clinical phenotype of muscular dystrophy, but the control method did not obtain diagnostic information. However, the method of this invention detected 2 cases of inversion variants, 8 of which were SNV / INDEL variants, 2 of which were DUP variants, and 2 of which did not show DMD variants. Sanger sequencing was performed on the samples with inversion variants to verify the breakpoints, and the Sanger sequencing results were consistent with the results detected by this invention. Of the samples that tested positive for DMD variants using the control method, 5 samples showed deletion ranges inconsistent with those detected by this invention. Sanger sequencing was performed on these 5 samples to verify the breakpoints, and the Sanger sequencing results were consistent with those detected by this invention. One sample tested positive for a complex rearrangement variant. Based on the breakpoint analysis, three possible variants were inferred.

[0079] Table 10: Detection in clinical samples

[0080] Based on existing detection performance, this invention can correctly detect missing, duplicate, and inverted structural variations, such as samples 1 and 9 mentioned above. This invention can correctly detect complex rearrangements caused by structural variations, and can infer the specific rearrangement based on the generated breakpoints. Specific detection results for the samples are as follows: Figures 1-5 As shown. This invention has a higher mutation detection rate than other technologies, enabling accurate detection of clinically pathogenic sites. This invention can detect specific sites of SV or SNV mutations, and based on these specific sites, reading frame change analysis can be performed, thereby predicting pathogenic mechanisms and phenotypes, providing information for protein function prediction, and offering more detailed reference information for clinical diagnosis.

[0081] For SNV variant sites, if the pathogenicity rating is not clear (VUS), there is insufficient evidence to support a diagnosis. Following the library construction methods in Examples 1 and 2, sequencing and analysis were performed on six clinical peripheral blood samples. The results show that this invention can provide assessment and prediction of deep intron variants, and provides the probability and score of deep intron variants, which can assist in clinical diagnosis (Table 11).

[0082] Table 11: Deep Intron (DIV) Analysis of Samples for which DMD Diagnostic Information Was Not Obtained

[0083] Example 4: Amniotic fluid sample DMD Variation and differential diagnostic variation analysis First, the amniotic fluid DNA was fragmented using a fragmentation tube to extract 50 ng of DNA. The subsequent construction of the third-generation sequencing library followed the same procedure as in Example 1, except that a pre-amplification step was added between the "pre-library adapter ligation" and "hybridization of the pre-capture library" steps. The reaction system and procedure were consistent with those for "PCR amplification and enrichment of the capture product." After constructing the third-generation library, slide selection was performed before sequencing, referring to Example 2. The probe combinations used are shown in the rightmost column of Table 12, where probe combination A is probe group (1) + probe group (6). The results are shown in Table 12 below.

[0084] Table 12: Prenatal Amniotic Fluid Samples DMD Detection of variants

[0085] This invention enables the detection of complex DMD variants in amniotic fluid samples. Because amniotic fluid samples are relatively precious and generally have low total DNA content, this invention only requires 50 ng of DNA to perform third-generation sequencing analysis. Current analysis can detect SNV / INDEL variants and SV variants, with SV variants detected precisely at specific breakpoints. This invention is also applicable to the analysis of complex structural variants. This invention is of great significance for prenatal diagnosis.

Claims

1. A probe assembly comprising probes (1) and (6) of the following probe groups: (1) Targeting DMD probe sets of gene regions, wherein DMD Gene regions contain DMD Full-length gene; (2) Targeting DMD The probe set of gene regulatory regions, wherein DMD Gene regulatory regions include DMD The 20 kb region upstream of the 5'UTR and the 20 kb region downstream of the 3'UTR of the gene; (3) Targeting DMD The probe set that modifies the gene region, wherein DMD Modified gene regions include SPP1 Key gene loci rs28357094, rs11730582, rs11439060, and rs17524488. LTBP4 Key gene loci: rs2303729, rs1131620, rs1051303, rs710160, and rs10880. CD40 The key gene site rs1883832 ACTN3 The key gene locus rs1815739 DYNLT5 Key gene loci rs1060575 and rs3816989, THBS1 Key gene loci rs2725797 and rs2624259, and ADRB2 The key gene locus is rs1042713. (4) A probe set targeting a diagnostic gene region, wherein the diagnostic gene region comprises the full length of the following genes: LMNA, PABPN1, CAPN3, DMPK, DYSF, TTN, COL6A1, CNBP, LAMA2, GNE, EMD, SGCA, SGCG, SGCB, SGCD, DYNLT5, FKRP, ANO5, CRPPA, POMT2, FKTN, POMGNT1, POMT1, DAG1, GMPPB, POMK, STIM1 ; (5) A probe set targeting the mitochondrial gene region, wherein the mitochondrial gene region contains the full length of the following genes: ND1, ND2, ND3, ND4, ND4L, ND5, ND6, CYTB, COX1, COX2, COX3, ATP6, ATP8, MT-RNR1, MT-RNR2, MT-TA (Ala), MT-TR (Arg), MT-TN (Asn), MT-TD (Asp), MT-TC (Cys), MT-TE (Glu), MT-TQ (Gln), MT-TG (Gly), MT-TH (His), MT-TI (Ile), MT-TL1, MT-TL2, MT-TK (Lys), MT-TM (Met), MT-TF (Phe), MT-TP (Pro), MT-TS1, MT-TS2, MT-TT (Thr), MT-TW (Trp), MT-TY (Tyr), MT-TV (Val); (6) A probe set targeting the sex identification gene region, wherein the sex identification gene region includes the full length of the SRY gene and its 2 kb upstream region of the 5'UTR and its 2 kb downstream region of the 3'UTR.

2. The probe assembly according to claim 1, comprising probe groups (1), (2), (3) and (6).

3. The probe assembly according to claim 2, comprising probe groups (1), (2), (3), (4), (5) and (6).

4. The probe assembly according to any one of claims 1-3, wherein... DMD Gene regulatory regions include DMD The 100 kb region upstream of the 5'UTR and the 100 kb region downstream of the 3'UTR of the gene.

5. The probe assembly according to any one of claims 1-3, wherein... DMD Modified gene regions further include SPP1 Full-length gene LTBP4 Full-length gene CD40 Full-length gene ACTN3 Full-length gene DYNLT5 Full-length gene THBS1 Full-length gene and ADRB2 Full-length gene.

6. The probe assembly according to any one of claims 1-3, wherein... DMD Modified gene regions further include SPP1 Full-length gene and its 5'UTR and 3'UTR regions, LTBP4 Full-length gene and its 5'UTR and 3'UTR regions, CD40 Full-length gene and its 5'UTR and 3'UTR regions, ACTN3 Full-length gene and its 5'UTR and 3'UTR regions, DYNLT5 Full-length gene and its 5'UTR and 3'UTR regions, THBS1 Full-length gene and its 5'UTR and 3'UTR regions and ADRB2 Full-length gene and its 5'UTR and 3'UTR regions.

7. The probe assembly according to any one of claims 1-3, wherein each probe satisfies the following requirements: (a) Target PABPN1, CAPN3, DMPK The full-length probes cover 1-3 kb regions of the 5'UTR and 3'UTR, and are designed with a 1× tiled coverage method; the remaining probes uniformly cover the target region and the interval between each two adjacent probes is 400-1000 bp. (b) No probes will be designed for areas where probes are not suitable; (c) The probe length is 80-120 bp and the GC content is 40%-60%.

8. The probe combination according to any one of claims 1-3, wherein each probe is a DNA probe or an RNA probe.

9. The probe assembly according to any one of claims 1-3, wherein each probe is connected to a label selected from at least one or more of biotin, avidin, streptavidin, antibodies, or chemical conjugates.

10. The probe assembly according to any one of claims 1-3, wherein each probe assembly comprises probes of one, more, or all of the following coverage types: (i) A probe that covers the entire length; (ii) Probes covering exon regions; (iii) Probes covering mutation hotspot regions; (iv) Probes covering key SNP sites.

11. A kit comprising the probe combination according to any one of claims 1-10.

12. The kit according to claim 11, further comprising: pre-capture library construction reagent; hybridization capture library construction reagent; post-capture PCR amplification reagent; and third-generation library construction reagent.

13. The kit of claim 12, wherein the pre-capture library construction reagent comprises a repair enzyme, a repair buffer, a ligase, a ligation buffer, and a adapter.

14. The kit according to claim 13, wherein the connector is a Y-type connector, a sticky end connector, or a flat end connector.

15. The kit of claim 12, wherein the hybridization capture library construction reagent comprises hybridization buffer, hybridization enhancement buffer, human cot-1 DNA, capture magnetic beads, capture washing reagent and optional sequence blocking reagent.

16. The kit of claim 15, wherein the sequence blocking reagent comprises a sequence that is inversely complementary to the sequence of the adapter.

17. The kit according to claim 12, wherein the post-capture PCR amplification reagent comprises an amplification enzyme, an amplification buffer, a PCR additive, and amplification primers.

18. The kit according to claim 17, wherein the PCR additive is selected from at least one of dimethyl sulfoxide, betaine, bovine serum albumin, and formamide.

19. The kit according to claim 17, wherein the amplification primers are phosphorylated.

20. The kit according to claim 12, wherein the third-generation library construction reagent comprises a repair enzyme, a repair buffer, a ligase, a ligation buffer, an exonuclease, an exonuclease buffer, a third-generation adapter, and a slice select reagent.

21. A method for constructing single-molecule long-read sequencing libraries, comprising the following steps: S1) Extract DNA from the sample to be tested and fragment it; S2) End repair and adapter ligation are performed on fragmented DNA to obtain a pre-capture library; S3) Hybridize the pre-capture library with the probes of any one of claims 1-10 for capture and PCR amplification to obtain the post-capture PCR product; S4) The captured PCR products are used to construct a third-generation sequencing library to obtain a single-molecule long-read sequencing library.

22. The method of claim 21, wherein the sample to be tested includes blood, dried blood spots, amniotic fluid, cultured amniotic fluid cells, hair, saliva, tissue, semen, sperm stains, and bone.

23. The method of claim 21, wherein the fragmentation comprises ultrasonic treatment, mechanical breaking, or enzymatic digestion.

24. The method of claim 21, wherein the length of the fragmented DNA is 7-15 kb.

25. The method of claim 21, wherein after end repair and adapter ligation in step S2, PCR amplification is further included.

26. The method of claim 21, wherein step S3 includes adding a closing sequence, the closing sequence being inversely complementary to the connector sequence.

27. The method according to claim 21, further comprising S5) slice selection of the single-molecule long-read sequencing library.

28. Used for detection DMD Gene, DMD Devices for modifying genes and identifying and diagnosing genes, including: 1) Pre-capture library construction module: used to fragment the DNA of the sample to be tested and obtain the pre-capture library; 2) Hybridization capture module: used to hybridize and capture the pre-capture library with the probes according to any one of claims 1-10 and then amplify it; 3) Third-generation sequencing library construction module: used to construct single-molecule long-read sequencing libraries; 4) Sequencing and analysis module: used for sequencing and analyzing variant information on a third-generation sequencing platform.

29. The device of claim 28, wherein the third-generation sequencing platform is selected from nanopore single-molecule sequencing platforms and single-molecule real-time sequencing platforms.

30. The device of claim 28, wherein the analysis of variation information includes the following process: i) The sequencing data of the samples to be tested are split and compared with the human reference genome for analysis. ii) Based on the compared data, analyze the relevant variants, including SNV, Indel, and SV, and annotate the variants to obtain the pathogenicity rating of the variants.

31. The device of claim 30, wherein the human reference genome is human reference genome version GRCh38.

32. The device according to claim 28, wherein the following can be achieved through the process defined in claim 29: a) Analyze and annotate the SNV variants in exons; b) Analyze the SNV variation of introns and use it for variation analysis, prediction, and annotation of deep introns; c) Analyze and detect breakpoint locations and scattered duplicate samples of DMD; d) Analyze and detect breakpoint locations and complex rearranged samples.

33. The use of the probe combination according to any one of claims 1-10 in the preparation of a kit for the diagnosis of Duchenne muscular dystrophy, detection of differential gene variants, differential diagnosis of muscular dystrophy, detection of DMD-related modified gene variants, and detection of DMD transcriptional regulatory region variants.

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