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Rapid and sensitive genotype identification and nucleic acid detection

A nucleic acid molecule and oligonucleotide probe technology, which is used in the fields of rapid and sensitive genotype identification and nucleic acid detection, and can solve the problems of limited membrane area and long incubation time.

Inactive Publication Date: 2016-03-09
DIAGCOR LIFE SCI LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional hybridization techniques are in principle limited by the membrane area and usually require long incubation times

Method used

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  • Rapid and sensitive genotype identification and nucleic acid detection
  • Rapid and sensitive genotype identification and nucleic acid detection
  • Rapid and sensitive genotype identification and nucleic acid detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] test program

[0075] The separation steps for DNA, PCR amplification, hybridization, detection samples and result analysis, such as WO / 2011 / 139750 or related procedures and techniques known to those skilled in the art can be applied to the present invention. Various internal controls (IC) can be included in the test test to verify the DNA sample, PCR amplification and color development of the results. The size and format of each guide film array can be used according to individual needs, or used to optimize the use of the reaction chamber to maximize cost savings.

Embodiment 2

[0077] Genotyping MDR-TB

[0078] In one embodiment, the present invention provides methods and kits for DR-MTB genotyping. The method and kit use polymerase chain reaction (PCR) and "drainage" hybridization technology to detect Mycobacterium tuberculosis (MTB) resistant to rifampicin (RIF) and isoniazid (INH) . The corresponding primers used to amplify the rpoB gene, katG gene and INHA gene have been confirmed not to cross-react with the human genome. RS-IAC is an internal control primer that is not targeted at the human or MTB genome and is used to monitor the PCR amplification process. The gene mutations used to detect drug resistance include rpoB gene (D516V, D516G, H526D, H526Y, H526L1, S531L and S531W), katG gene (S315T1 and S315T2), and inhA gene (-15TC / T). There are also five probes for detecting wild-type MTB mutations, which helps to verify whether the PCR amplification reaction of rpoB gene, katG gene and inhA gene has been completed. In one embodiment, the primer...

Embodiment 3

[0097] Beta Mediterranean Genotyping

[0098] The present invention provides a systematic detection of β-globulin gene mutations. The gene mutations include TATA-28 (A> G), TATA-29 (A> G), start codon (G> A), detection of codon 5 (-CT), codon 8 / 9 (+G), codon 15 (G> A), codon 16 (-C), codon 17 (A> T), codon 19 (A> G), codon 26 (G> A) (hemoglobin E), codon 27 / 28 (+ three), codon 30G> C, IVS1.1 (G> T), IVS1.1 (G> A), IVS1.5 (G> C), codon 41 / 42 (-TCTT), codon 43 (G> T), codon 71 / 72 (+A), IVS2.1 (G> A), IVS2.654(C> T) and a deletion of 619 bp.

[0099] The present invention also provides a method for detecting β-globulin mutations in a sample of a subject, the method comprising the following steps: (a) obtaining a sample containing template nucleic acid molecules from the subject; (b) with a sequence number selected from : The set of 201-205 primers amplifies the template nucleic acid molecule, thereby generating an amplified product; and (c) the amplified product is composed of an olig...

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Abstract

The invention discloses a method, primer, probe and kit for identifying various gene mutations or substance genetic typing. In one embodiment, the disease is pathogenic. In the other embodiment, the invention is applied to detection of multidrug-resistant Mycobacterium tuberculosis, hepatitis B virus, beta-globulin mutation, thrombophilia related mutation; or various sexually transmitted pathogens, consisting of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum, Treponema pallidum, Herpes simplex virus 1 , Herpes simplex virus 2, Human papillomavirus type 6, and Human papillomavirus type 1.

Description

[0001] Related application [0002] This application claims priority based on the US Provisional Patent Application No. 61 / 791,933 filed on March 15, 2013, and all contents of the patent application are incorporated into this application by reference. Technical field [0003] The field of the present invention is to identify various genotypes related to human diseases. Background technique [0004] The invention relates to the identification of different genotypes. Sequence-specific primer-polymerase chain reaction (SSP-PCR), DNA sequencing, DNA fingerprinting, and single nucleotide polymorphism (SNP) genotyping techniques have been applied to genotyping (PCT application publication No. WO / 2011 / 139750). Among these techniques, single nucleotide polymorphism (SNP) genotyping has a higher recognition power. However, most of the genotyping tests involving DNA hybridization require higher running costs and longer operation time. Therefore, it is necessary to develop more effective ge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6816C12Q2531/113C12Q1/68C12Q1/6883C12Q1/689C12Q1/703C12Q2600/106C12Q2600/156A61B8/5223A61B8/08A61B8/4236
Inventor 谭约瑟夫荣安周约瑟夫国辉郭秀梅杨温迪颖珊朱丽安
Owner DIAGCOR LIFE SCI LTD
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