Gene chip for typing mycobacteria and application method thereof

A mycobacteria and gene chip technology, applied in the field of gene chip detection, can solve the problems of low sensitivity, high false positive rate, lack of unified operating standards and judgment standards, etc., and achieve the effect of high sensitivity and strong specificity

Inactive Publication Date: 2015-12-02
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these molecular biology methods have certain advantages for the identification of mycobacteria, they all have certain defects, such as the lack of unified operating standa...

Method used

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  • Gene chip for typing mycobacteria and application method thereof
  • Gene chip for typing mycobacteria and application method thereof
  • Gene chip for typing mycobacteria and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Design and development of gene chip for mycobacterial typing detection

[0051] 1. Target strain acquisition and DNA extraction

[0052] Purchasing 23 ATCC standard strains, namely Mycobacterium intracellulare (ATCC13950), Mycobacterium phlei (ATCC11758), Mycobacterium xenopiSchwabacher (ATCC19250), Mycobacterium smegmatis (ATCC19420), Mycobacterium triviale Kubica, ATCC23292, Mycobacterium terrae Wayne, ATCC15755, Mycobacterium gordonae, ATCC14470, Mycobacterium chelonae, ATCC35752, Mycobacterium tuberculosis, ATCC , Mycobacterium aurum Tsukamura, ATCC23366, Mycobacterium ulcerans, ATCC19423, Mycobacterium malmoense, ATCC29571, Mycobacterium avium, ATCC25291, Mycobacterium abscessus, ATCC19977), Mycobacterium fortuitum (ATCC6841), Mycobacterium scrofulaceum (ATCC19981), Mycobacterium gilvum (ATCC43909), Mycobacterium shimoidei Tsukamura (ATCC27962), Mycobacterium shimoidei Tsukamura, ATCC27962 (Mycobacterium gastriWayne, ATCC15754), Mycobacterium nonchromo...

Embodiment 2

[0097] Example 2: Determination of Positive Judgment Criteria for Mycobacterium Typing Detection Gene Chip

[0098] The cutoff value is a criterion for judging whether the signal value of the gene chip is positive. For each typing probe, non-target strains (i.e. negative strains) and blank controls were selected for gene chip hybridization. Through repeated experiments and data statistics, the background statistical average value of negative strains and blank controls + 2SD was used as each probe The Cutoff value. The discrimination ability of each mutation detection probe was more than 2.5 times as the positive judgment standard.

Embodiment 3

[0099] Example 3: Specificity Evaluation of Gene Chip for Mycobacterium Typing Detection

[0100] Specificity is the most important assessment index in diagnosis. The gene chip of the present invention uses the optimized system and conditions to detect the specificity among all target strains. See the appendix of the manual for some test results. image 3 , it can be seen that all can be clearly distinguished, indicating that the specificity of the present invention is good.

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Abstract

The invention discloses a gene chip for typing mycobacteria and an application method thereof and relates to the technical field of gene chip detection. The application method includes: 1, designing a mycobacteria typing specific probe; 2, preparing the chip; 3, extracting and amplifying nucleic acid; 4, hybridizing the chip; 5, developing and analyzing the chip. The species specific probe is designed by analyzing 23 common mycobacteria hsp65 gene sequences, a genome DNA of a sample to be detected is used as a template, a marked primer group is used for amplifying, an amplified product is hybridized with the probe on a bio-chip, and types of mycobacteria are determined according to hybridizing results. The gene chip and the application method thereof have the advantages such as high specificity, high sensitivity, results stability and reliability and fitness for batch sample detection.

Description

Technical field: [0001] The invention relates to a gene chip for typing mycobacteria and a method for using the same, belonging to the technical field of gene chip detection. Background technique: [0002] Tuberculosis caused by Mycobacterium tuberculosis infection is one of the main infectious diseases in humans today. Since 1985, the incidence of tuberculosis has gradually increased worldwide. A report from WHO showed that the incidence of tuberculosis was 143 cases per 100,000 people in 1990, and rose to 152 cases per 100,000 people in 1995. 162 cases per 100,000 people. According to statistics, about one-third of the world's population has been infected with Mycobacterium tuberculosis. Every year, 8 million people are infected with tuberculosis, of which about 3 million people die. Tuberculosis has become a global public health problem. my country is the second largest high-risk country for tuberculosis in the world. The number of tuberculosis patients accounts for 70%...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C40B40/06C12R1/32
CPCC12Q1/689C12Q1/6837C12Q2600/158C12Q2600/16C40B40/06C12Q2537/143C12Q2565/501
Inventor 宋玉竹夏雪山巩振立赵勤李晓霏张阿梅李道群杨雪枫
Owner KUNMING UNIV OF SCI & TECH
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