Method and kit for rapidly detecting single nucleotide polymorphisms (SNPs)

A technology for single nucleotide polymorphism and detection method, which is applied in the field of rapid detection of single nucleotide polymorphism, and can solve the problems of strong professionalism, complicated operation and high price.

Active Publication Date: 2012-08-01
USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of the various methods currently on the market require special analytical instruments, which are not only expensive, but also complicated to operate and highly professional, so they are limited in application.

Method used

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  • Method and kit for rapidly detecting single nucleotide polymorphisms (SNPs)
  • Method and kit for rapidly detecting single nucleotide polymorphisms (SNPs)
  • Method and kit for rapidly detecting single nucleotide polymorphisms (SNPs)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Example 1 Detection and typing of mitochondrial DNA G11778A single nucleotide polymorphism in Leber's disease

[0106] Leber's hereditary optic neuropathy (Leber's hereditary optic neuropathy, LHON) is a maternally inherited binocular optic nerve disease. In 1988, Wallace et al. first reported that the primary mutation (G→A) at the 11778th nucleotide site of mitochondrial DNA (mtDNA) could cause LHON. So far, 25 mtDNA site mutations related to this disease have been found, so molecular biological genetic examination has become the preferred method to identify Leber's disease of different etiologies. In view of the diagnostic significance of mitochondrial G11778A site typing for Leber's hereditary optic neuropathy, the SNPS test strip newly invented by this patent is used to detect the polymorphism of G11778A site. The specific implementation is as follows:

[0107] (1) Sampling

[0108] The subject's peripheral venous anticoagulant blood (frozen storage) was 5 microl...

Embodiment 2

[0161] Example 2. Detection of Mycobacterium tuberculosis isoniazid resistance mutation KatG 315 mutation

[0162] Isoniazid is an important drug for the treatment of Mycobacterium tuberculosis, but long-term use will produce drug resistance. Mycobacterium tuberculosis-specific gene mutations are the primary cause of resistance to isoniazid, and the mutations are mainly AGC→AAC and AGC→ACC at position 315 of the Kat G gene. Therefore, dynamic monitoring of the KatG 315 mutation of Mycobacterium tuberculosis has important guiding significance for timely adjustment of treatment regimens, rational use of drugs and improvement of efficacy.

[0163] (1) Sampling

[0164] The DNA extraction kit produced by Hangzhou Ustar Biotechnology Co., Ltd. was used to extract nucleic acid from the sputum of patients with Mycobacterium tuberculosis for gene amplification.

[0165] (2) PCR-AS-PCR nucleic acid test strip method to detect the KatG 315 mutation of mycobacteria:

[0166] Non-speci...

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PUM

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Abstract

The invention relates to an SNPs detection method, and concretely relates to a method for detecting specifically amplified products through a one-step PCR reaction and a nucleic acid detection test paper strip in a same reaction system. The method comprises the following steps: nonspecifically amplifying fragments containing SNPs sites through first PCR temperature cycling; specifically amplifying bases containing the SNPs sites through AS-PCR (allele specific polymerase chain reaction); and detecting the specifically amplified products by the nucleic acid detection test paper strip. The method can also be used for detecting mononucleotide mutation. The method which has the characteristics of high sensitivity, strong specificity, simple operation, short time, and rapid and reliable detection result combines respective advantages of proteins and nucleic acids. The invention also relates to applications of a kit in the detection of known drug-resistant gene of pathogens, the mating selection of a donor and an acceptor in organ transplantation, and the discrimination of the identity of a criminal or paternity tests in forensic researches.

Description

technical field [0001] The present invention relates to a kind of detection method of single nucleotide polymorphism, specifically, it relates to a kind of PCR amplification technology and nucleic acid test strip detection technology which are completed by two cycle programs of non-specific amplification cycle and specific detection cycle A technique for the rapid detection of single nucleotide polymorphisms. Background technique [0002] SNPs refer to the existence of two or more different bases at a specific nucleotide position at the genome level, and the frequency of any allele in the population is not less than 1%. SNPs have become the third generation of molecular genetic markers following restriction fragment length polymorphism (Restriction Fragment Length Polymorphism, RFLP) and short tandem repeat (Short Tandem Repeat, STR) polymorphic markers. Therefore, the detection of SNPs is becoming the focus of widespread attention. At present, most of the methods for SNPs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 胡林徐高连尤其敏王宏莹高谦张文宏梅建
Owner USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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