112 results about "Beta thalassemia" patented technology

Disclosed herein are methods and low dose regimens for increasing fetal hemoglobin levels in patients with red blood cell disorders, such as beta thalassemia, sickle cell disease, other anemias, or blood loss. Fetal and total hemoglobin levels and red blood cell counts are increased by administering 2,2-dimethylbutyrate (DMB) alone or in combination with hydroxyurea, decitabine or an HDAC inhibitor. Treatment can be continued for at least two weeks.

The invention provides primers, a method and a chip for detecting alpha and/or beta-thalassemia point mutation and deletion mutation based on whole-gene capture sequencing and application of such primers, such method and such chip. The primers, the method, the chip and application thereof have the advantages that through designing of capture probes, relevant genes involved in alpha-thalassemia and beta-thalassemia are enriched and all mutation information including SNP and indel in full-length sequences of genes is detected; through addition of autosome, X-chromosome and Y-chromosome regions as well as upstream and downstream regions of coded genes as references, structure variations such as SNV and CNV are detected; compared with existing various hotspot mutation site detection technologies, the method is capable of detecting hotspot mutation information as well as some rare mutations and undiscovered new mutation types to detect and analyze full-length sequence specificity of target genes, fully covers the mutation types and makes up the defect that a conventional detection method easily causes missing detection of low-frequency mutations and rare mutations greatly.

The invention belongs to the technical field of biology and particularly relates to a primer set and a kit for detecting rare deletion type thalassemia. The primer set and the kit can be directly used for rapidly and stably detecting domestic 11 types of known rare deletion type thalassemia. The primer set for detecting the rare deletion type thalassemia comprises a primer set A and a primer set B, wherein the primer set A comprises 12 primers for detecting alpha-thalassemia deletion gene types; the primer set B comprises 10 primers for detecting beta-thalassemia deletion gene types. The kit can adopt a multiplex Gap-PCR (Polymerase Chain Reaction) technology to detect 11 rare deletion gene types and can be used for directly detecting 11 rare deletion thalassemia gene types for one time. The kit is simple to operate and saves time and cost when being compared with a manner of combining a nested PCR with a genetic analysis in antenatal diagnosis; conditions are created for comprehensively carrying out thalassemia screening and scientific evidences are provided for thalassemia diagnosis of premarital detection, antenatal detection and fetuses of the pregnancy.

The invention discloses a human beta globin gene containing a human beta globin gene promoter, a recombinant adeno-associated virus vector containing the human beta globin gene, and a method for preparing the recombinant vector. The recombinant adeno-associated virus vector inserts an HS2 segment, an HS3 segment and an HS4 segment of a human beta globin gene cluster enhancer core sequence and a human beta globin gene sequence containing the human beta globin gene promoter into repeated sequence ITRs on the reverse terminal of an adeno-associated virus. The recombinant vector has high transfection efficiency; and mediated exogenous genes can be expressed in vivo for a long time, have good safety and can be used for gene therapy of beta Mediterranean anemia.

The invention relates to a fluorescence quantitative PCR detection kit for beta-thalassemia mutation. The kit comprises a PCR mixing reaction liquid, a positive control and fluorescence probes for detecting beta-thalassemia mutation genotype, wherein the PCR mixing reaction liquid contains PCR primers for amplifying a gene fragment on which a mutation site is positioned, and the mutation site is at least one mutation site selected from deletion mutation of a base corresponding to the site 41/42 amino acid of beta-globin gene, C-to-T mutation of a base corresponding to the site 654 amino acid of the second intron of beta-globin gene, A-to-T mutation of a base corresponding to the site 17 amino acid of beta-globin gene, A-to-G mutation of a base corresponding to the site 28 amino acid on the upstream of the promoter of beta-globin gene, A base insertion mutation of a base corresponding to the site 71/72 amino acid of beta-globin gene, and G-to-C mutation of a base corresponding to the site 5 amino acid of the first intron of beta-globin gene. With the technical scheme of the present invention, rapid, accurate and high sensitivity detection of mutation conditions of beta-thalassemia gene can be achieved, and especially 6 special site mutations of beta-thalassemia gene can be detected, wherein the 6 special site mutations are common in Chinese.

The invention relates to a kit and particularly relates to a kit for editing or repairing human hemoglobin gene. The kit comprises sgRNA of specific target HBB gene or a carrier containing the nucleotide sequence of the sgRNA; and/or modified Cas9 enzyme or a sequence encoding the modified Cas9 enzyme or a carrier containing the sequence encoding the modified Cas9 enzyme; and/or a donor gene sequence for repairing the HBB gene of beta-thalassemia or a carrier containing the donor gene sequence. The invention also provides an application of the kit in cutting or repairing the HBB gene, especially an application in repairing the HBB gene of autologous hematopoietic stem cells of a patient with beta-thalassemia. The kit provided by the invention has the advantages that the targeting capability of the target HBB gene is strong, the cutting efficiency of the HBB gene is high, the off-target efficiency is low, the HBB gene can be effectively repaired and the prospect in clinical study and treatment application is wide.

Methods of using one or more fumaric acid esters or pharmacologically active salts, derivatives, analogues, or prodrugs thereof to increase expression of fetal hemoglobin (HbF) are disclosed. The methods typically include administering to a subject an effective amount of one or more fumaric acid esters optionally in combination or alternation with hydroxyurea to induce HbF expression in the subject in an effective amount to reduce one or more symptoms of a sickle cell disorder, a hemoglobinopathy, or a beta-thalassemia, or to compensate for a genetic mutation is the human beta-globin gene (HBB) or an expression control sequence thereof. Pharmaceutical dosage units and dosage regimes for use in the disclosed methods are also provided.

The invention relates to a beta-thalassemia mutation detection kit, in particular to a kit for detecting the nucleotide polymorphism of beta-thalassemia by an asymmetric amplification method. The kit comprises a set of specific nucleotide polymorphism probes and a polymerase chain reaction (PCR) primer for amplifying a target gene in a sample, and can detect 27 types of beta-thalassemia mutations; and the detection process is quicker, and the detection result is more accurate.

The invention relates to a method and kit for detecting alpha and beta thalassemia point mutation based on a next generation sequencing technology .In the detection method, PCR (polymerase chain reaction) amplification primer sequences SEQ ID NO1-17 in exon, regulation and transcription regions of HBA1, HBA2 and HBB genes, PCR marker primer sequences SEQ ID NO18-115 at the 5'-terminal and PCR marker primer sequences SEQ ID NO116-213 at the 3'-terminal involved with alpha and beta thalassemia are included. The detection method comprises the following detection steps: (1) constructing a next generation sequencing library; (2) purifying the next generation sequencing library; (3) sequencing through a next generation sequencer; and (4) performing bioinformatic analysis to obtain a result. The kit for detection comprises the above-mentioned primer sequences and the next generation sequencer solexa. The detection method provided by the invention has the characteristics of simple operation steps, high detection specificity, wide detection range, low cost and the like.

Disclosed are a thalassemia mutant gene detection primer pair and kit, and application and library construction methods thereof. The thalassemia mutant gene detection primer pair comprises primer sequences shown as SEQ ID NO: 1-8. The thalassemia mutant gene detection primer pair can be applied to simultaneously detecting a number of mutation types and help discover new mutation types, and when combined with second-generation sequencing, can greatly reduce the detecting cost and improve throughput, thereby achieving significant cost advantages and being beneficial to popularized application ofthalassemia mutant gene detection.

The invention provides a method for detecting HBB gene mutation and HLA genotyping based on the high throughput sequencing technology and a corresponding reagent kit. An adopted primer composition comprises a primer of closely-linked single nucleotide polymorphisms (SNP) within the 1 Mb range of the up stream and the down stream of the specific amplification human embryo beta-thalassemia HBB gene and primers of the closely-linked single nucleotide polymorphisms (SNP) within the ranges at the up stream of the LHA-A gene, between the HLA-A gene and the HLA-B gene, between the HLA-B gene and the HLA-DRA gene, between the HLA-DRA gene and the HLA-DQB1 gene and at the downstream of the HLA-DQB1 gene of the specific amplification human leucocyte antigen system. The method has the advantages of university, single nucleotide polymorphisms (SNP) sequencing, high throughput, low cost, high flexibility and strong specificity.

The invention belongs to the field of biological chemical detection and particularly relates to a method of determining the ratio of a globin chain alpha to a globin chain beta in hemoglobin and application of the method in detection of beta-mediterranean anemia. The method comprises the steps that hemoglobin samples are taken and split to obtain split fragments of the globin chain alpha and split fragments of the globin chain beta; one split fragment of the globin chain alpha and one split fragment of the globin chain beta are selected and used as sign peptide fragments; a mass spectrum is used for determining the concentration of the sign peptide fragment of the globin chain alpha and the concentration of the sign peptide fragment of the globin chain beta; the ratio of the globin chain alpha to the globin chain beta is expressed just by the ratio of the concentration of the sign peptide fragment of the globin chain alpha to the concentration of the sign peptide fragment of the globin chain beta. According to the method, by means of quantitative analysis of the ratio of the globin chain alpha to the globin chain beta, different types of beta-mediterranean anemia can be identified at an early stage, and severe, medium and mild beta-mediterranean anemia can be identified.

The invention discloses an HBB gene kit for correcting autologous hematopoietic stem cells of a patient suffering from servious beta-thalassemia. The kit consists of a set of reagents for preparing a specific HBB-101 HIV slow virus and a set of reagents for HIV slow virus infection hematopoietic stem cells. The invention further discloses application of the kit in converting the autologous hematopoietic stem cells of the patient suffering from servious beta-thalassemia into hematopoietic stem cells with normal beta-globin synthesis functions. Experiment shows that the kit disclosed by the invention is simple to operate and relatively high in virus titer, and has wide development prospects in clinical study and treatment application, the virus infection efficiency is as high as 56.99%, and the purpose of treating the patient suffering from servious beta-thalassemia can be achieved through venous re-transfusion after PCR identification on corrected hematopoietic stem cells is implemented and the result of DNA sequencing identification is positive.

The present invention provides a primer set for detecting alpha thalassemia and beta thalassemia, and belongs to the field of molecular biology, wherein the primer set comprises an alpha primer set and a beta primer set. The invention further provides a chip capable of simultaneously detecting alpha thalassemia and beta thalassemia, wherein probes having sequences represented by SEQ ID NO:1-31 areimmobilized on the chip. Based on the primer set and the chip, the present invention further provides a kit capable of simultaneously detecting alpha thalassemia and beta thalassemia. The kit of theinvention has advantages of short detection time, low cost, convenient operation, closed tube operation and pollution reducing, and has great significance in the screening of thalassemia population, the genetic counseling and the prenatal diagnosis.

The invention discloses a noninvasive antepartum fetal beta-thalassemia gene mutation detection library building method, a detection method and a kit. In the library building method, a specific connector is connected onto a maternal peripheral blood free DNA (deoxyribonucleic acid) fragment; then, a connector connection product pre-amplification product is divided into two parts; two rounds of specific amplification are performed by respectively and independently using forward primers and reverse primers aiming at target sites; the target sites can be enriched at high specificity; the amplification specificity of the primers can be obviously improved. The forward and reverse primer groups of a plurality of SNP (single nucleotide polymorphism) sites used for calculating the fetal free DNA proportion are respectively used for performing two-round specific amplification; the fetal free DNA proportion can be efficiently and accurately calculated. The library is subjected to sequence testing; the beta-thalassemia gene mutation can be accurately and effectively detected; the result is identical to the result of the amniocentesis detection and typing; the safety, the noninvasion and the high efficiency are obviously superior to those of the amniocentesis detection.

The invention discloses a virus vector for expressing recombinant human beta-globin and application of the virus vector. The invention provides a DNA fragment A which is formed through linking of thefollowing components from a 5' terminal to a 3' terminal: three DNA enzyme I hypersensitive sites, an HBB promoter, a translation enhancer, an HBB gene expression frame and an HBB transcription enhancer. An HBB expression vector provided by the invention has the capability of efficiently expressing exogenous HBB proteins and has extremely high application values in treatment of beta-thalassemia and sickle cell anemia.

The invention relates to a nucleic acid composition and a detection kit for detecting genetic anemia as well as a use method. The nucleic acid composition and the detection kit for detecting the genetic anemia can be used for simultaneously detecting four deletion type alpha thalassemia genes, three non-deletion type alpha thalassemia genes, nineteenth mutation type beta thalassemia genes, geneticanemia types such as sickle-shaped thalassemia gene as well as specific gene mutation types. Compared with the prior similar technologies, the detection kit for the genetic anemia has the characteristics that several mutation detection types for the genetic anemia are increased, such as alphaTHAI deleted thalassemia, sickle-shaped thalassemia and 71/72(+T) mutation and -28M(A-C) mutation which are relatively-rare thalassemia types. The detection for the genetic anemia types can provide visual reference and prompt for clinically detecting the genetic anemia, so that the leak detection risk ofclinical genetic anemia can be greatly reduced and the birth rate of severe anemia children is reduced.

The invention discloses a beta-thalassemia gene amplification kit and a method based on a recombinase polymerase amplification technology. The kit comprises RPA amplification primers which are respectively P1F-1 shown as SEQ ID NO. 1, P1Rbio-1 shown as SEQ ID NO. 2, P2F-1 shown as SEQ ID NO. 3, P2Rbio-1 shown as SEQ ID NO. 4, P3F-1 shown as SEQ ID NO. 5 and P3Rbio-1 shown as SEQ ID NO. 6. The amplification is performed on genomic DNA of a to-be-detected specimen by using the RPA amplification primers and can be completed within 30 minutes, and the sensitivity is high; an obtained amplificationproduct can be detected by a liquid phase chip, and the accuracy rate is high.

The invention provides a probe and kit for detecting fetal free DNA beta-thalassemia mutation in maternal blood. The kit comprises a detection probe, a capture probe, a reaction film strip, a target gene amplification reagent, a cleaning solution, an incubation solution and a developing solution. According to the kit, a dot blot hybridization method is utilized; in the method, a DNA hybridizationprocess is used for detecting a product; and the kit has the advantages of simplicity in operation, rapidness, simplicity, convenience, small sample dosage, intuitive and reliable result and the like,and can be widely applied to detection of fetal free DNA beta-thalassemia mutation in maternal plasma in noninvasive prenatal diagnosis.

The invention provides a probe composition and a kit for screening genetic disease pathogenic gene carriers and a preparation method of the probe composition and the kit. The probe composition specifically captures pathogenic genes of genetic diseases, and the genetic diseases comprise alpha-thalassemia, beta-thalassemia, spinal muscular atrophy, hepatolenticular degeneration and methylmalonic acid cblC type. The invention provides a probe combination which is accurate, reliable, simple and economical, can be used for screening carriers of various genetic disease pathogenic genes at the same time, and is convenient for popularization and clinical application of screening of carriers of common genetic diseases.

The invention discloses a kit for rapidly detecting five beta-thalassemia mutations and application thereof, and belongs to the field of disease detection products. The kit comprises an amplification primer pair of five common mutation sites of beta-thalassemia genes and also comprises 2xconc.Master Mix and magnesium ion. The application comprises the following steps: performing polymerase chain reaction (PCR) amplification on an analysis sample by using the kit, performing high resolution melting curve analysis, and comparing with a positive melting curve of the five mutation sites so as to determine whether mutations and mutation types exist in the five sites. The kit can simultaneously detect one or combination of more than one in the five mutation sites, comprising -28, CD17, CDs41/42, CDs71/72 and IVS-II-654, of the beta-thalassemia mutation, specific probes and sequencing are not needed, the needle sample amount is small, and the kit is rapid in detection, accurate and low in cost and is suitable for large-scale detection.