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Primer for identifying mycobacterium tuberculosis complex strains and application of primer

A technology of mycobacterium tuberculosis and tuberculosis branches, which is applied in the field of medical inspection and can solve the problems of false positives, low resolution, and high price in image results

Active Publication Date: 2015-05-27
BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

IS6110-RFLP is widely used and has high resolution, but this method is based on the cultivation of a large number of strains, which is expensive, takes a long time, and strains with a small number of copies of IS6110 are not easy to type; MIRU-VNTR does not require a large number of cultured strains, and the results are easy to read , but due to the different locus combinations, the resolution and clustering rate are also different; the Spoligotyping method takes less time and costs less, but the resolution is lower, and the picture results are prone to false positives; SNP needs to be sequenced, and the price Expensive, not scalable across labs

Method used

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  • Primer for identifying mycobacterium tuberculosis complex strains and application of primer
  • Primer for identifying mycobacterium tuberculosis complex strains and application of primer
  • Primer for identifying mycobacterium tuberculosis complex strains and application of primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1. Primer pairs and identification methods for identifying Mycobacterium tuberculosis complex strains

[0078] 1. Primer pairs used to identify the Mycobacterium tuberculosis complex

[0079] The present invention designs PCR primer pair 1-primer pair 7 respectively for 16S rRNA, Rv0577, IS1561, Rv1510, Rv1970, Rv3877 / 8 and Rv3120. The PCR product sizes of the 7 pairs of primers are 975bp, 588bp, 466bp, 802bp, 711bp, 1055bp respectively and 321bp. Primer sequences are shown in Table 1.

[0080] Table 1, 7 pairs of primer information

[0081]

[0082]

[0083] 2. A method for rapid identification of Mycobacterium tuberculosis complex strains

[0084] 1. PCR amplification

[0085] Genomic DNA of the strain to be tested was extracted, and the obtained genomic DNA was used as a template, and the 7 pairs of primers designed in step 1 were used for PCR amplification.

[0086] Reaction system (20 μl): 10 μl of 2×Premix Taq, 1 μl of 10 μM upstream and downstr...

Embodiment 2

[0103] Embodiment 2, the application of Mycobacterium tuberculosis complex strain primer

[0104] 1. Tested strains: Mycobacterium tuberculosis M.tuberculosis (ATCC 27294); Mycobacterium bovis M.bovis Karlson and Lessel (ATCC 19210); BCG M.bovis BCG (ATCC35737); Mycobacterium microti M.microti Reed (ATCC 19422); M. canetii, M. caprae, M. africanum Castets et al. (ATCC 35711); M. avium subsp.avium Chester (ATCC 25291).

[0105] 2. Using the method for rapidly identifying Mycobacterium tuberculosis complex species in Example 1 to verify the above strains to be tested.

[0106] 3. Verification result

[0107] The result is as Figure 1-Figure 8 shown.

[0108] From figure 1 It can be seen that 7 pairs of primers detected 7 bands in total on the PCR product of Mycobacterium tuberculosis M.tuberculosis (ATCC27294): 975bp was amplified by primer pair 1, 588bp was amplified by primer pair 2, and 588bp was amplified by primer pair 3 466bp was obtained, 802bp was amplified by pri...

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Abstract

The invention discloses a primer for rapidly identifying mycobacterium tuberculosis complex strains and an application of the primer. The primer for rapidly identifying the mycobacterium tuberculosis complex strains consists of a primer pair 1, a primer pair 2, a primer pair 3, a primer pair 4, a primer pair 5, a primer pair 6 and a primer pair 7. The experiment shows that the primer can be used for accurately identifying the mycobacterium tuberculosis complex strains, the 7 pairs of the primers are under one same amplification reaction condition, amplification of 7 different primers is completed at one time, multiple times of amplification is not needed, the operation is simplified, and the time can be saved as the operation is rapid to implement.

Description

technical field [0001] The invention belongs to the field of medical examination, and in particular relates to a primer for distinguishing mycobacterium tuberculosis complex species and its application. Background technique [0002] Tuberculosis is one of the most serious epidemic infectious diseases, which poses a huge challenge to human public health. It has a very high morbidity and mortality rate worldwide. Some species of the Mycobacterium tuberculosis complex (MTBC) are the pathogenic bacteria that cause tuberculosis in humans and animals. Mycobacterium tuberculosis (M.tuberculosis), the main pathogen that causes human tuberculosis, originated about 3 million years ago and continues to affect human health due to the emergence of drug-resistant strains. In addition to Mycobacterium tuberculosis, other strains of MTBC have been found in patients, including Mycobacterium bovis (M.bovis), Mycobacterium Kennedy (M.canetii), Mycobacterium africanum (M.africanum subtype I),...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/32
CPCC12Q1/689C12Q2600/16
Inventor 孙照刚黄海荣苗慧芳朱宝利陈素婷刘飞徐玉辉
Owner BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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