Rapid genotyping analysis and kits thereof

A technology of nucleic acid molecules and oligonucleotide probes, which is applied in the field of rapid and sensitive genotype identification and nucleic acid detection, and can solve problems such as limited membrane area and long incubation time

Active Publication Date: 2017-06-30
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 genotyping analysis and kits thereof
  • Rapid genotyping analysis and kits thereof
  • Rapid genotyping analysis and kits thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0075] test program

[0076] Separation steps for DNA, PCR amplification, hybridization, detection of 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 assay to verify DNA samples, PCR amplification and color development of results. The size and format of each baffle array can be tailored to individual needs, or to optimize chamber usage for maximum cost savings.

Embodiment 2

[0078] Genotyping MDR-TB

[0079] In one embodiment, the present invention provides methods and kits for genotyping DR-MTB. The method and the kit adopt polymerase chain reaction (PCR) and "flow-through" hybridization technology for detecting 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 does not target the human or MTB genome and is used to monitor the PCR amplification process. The probes used to detect drug-resistant gene mutations included the rpoB gene (D516V, D516G, H526D, H526Y, H526L1, S531L, and S531W), the katG gene (S315T1 and S315T2), and the inhA gene (-15TC / T). There are also five probes for the detection of mutations in wild-type MTB, which help to verify whether the PCR amplification reactions of the rpoB gene, katG gene, and inhA gene ...

Embodiment 3

[0094] Beta Med Genotyping

[0095] The present invention provides a systematic detection of β-globulin gene mutation, said gene mutation includes TATA-28 (A>G), TATA-29 (A>G), initiation 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 619bp deletion.

[0096] The present invention also provides a method for detecting β-globulin mutation in a subject sample, the method comprising the following steps: (a) obtaining a sample comprising a template nucleic acid molecule from the subject; A group consisting of primers of 201-205 amplifies the template nucleic acid molecule, thereby generating an amplification product; and (c) the amplification product is composed of an oligonucleotide probe selected from the sequence ...

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Abstract

The present invention provides methods, primers, probes, and kits for identifying gene mutation or material genotyping. In one embodiment, the materials are disease-causing agents. In another embodiment, the present invention is applied to detecting multidrug-resistant mycobacterium tuberculosis, hepatitis B virus, beta-globulin mutation, thrombosis-related mutations; or multiple sexually transmitted diseases causing agents, including but not limited to, trichomonas vaginalis, chlamydia, neisseria gonorrhoeae, mycoplasma, mycoplasma hominis, ureaplasma urealyticum, Mycelia, Treponema pallidum, Herpes simplex virus type 1 and type 2 and human papilloma virus type 6 and type 11.

Description

[0001] This application is the PCT international application PCT / CN2014 / 000275 filed on March 17, 2014, which entered the Chinese national phase on November 3, 2015. The application number is 201480027795.3 and the invention name is "Rapid and Sensitive Genotype Identification and Nucleic Acid Detection "A divisional application of the invention patent application. [0002] related application [0003] This application claims priority based on US Provisional Patent Application No. 61 / 791,933, filed March 15, 2013, the entire contents of which are incorporated herein by reference. technical field [0004] The field to which the present invention relates is the identification of various genotypes associated with human disease. Background technique [0005] The present invention relates to the identification of different genotypes. Techniques such as sequence-specific primer-polymerase chain reaction (SSP-PCR), DNA sequencing, DNA fingerprinting, and single nucleotide polymor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12Q1/04
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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