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Mycobacterium PCR-RLB inspection and its primer and probe

A detection method and technology for mycobacteria, which are applied in the fields of detection and identification of mycobacteria, detection of mycobacteria by PCR-RLB, primers and probes, and can solve the problems of inability to identify and unsuitable for mixed infection, etc.

Inactive Publication Date: 2007-01-17
深圳市慢性病防治院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

"Chinese Journal of Antituberculosis" 2002, 24 (Supplement) P12-16 entitled "Research on Identification of Mycobacterium Species by Oligonucleotide Probes of 16 DNA Transcribed Spacer Sequences" uses reverse dot hybridization technology to 16 rRNA can only identify 9 mycobacteria in SGM (avian, intracellular, scrofula, sulga, sea, toad, Gordon, soil and achromogenic mycobacteria) and 3 complex groups (mycobacterium tuberculosis Ulcers / maritime, M. Kansas, Stomach and Scrofula complexes), 11 SGMs and M. ulcers / maritime, M. tuberculosis complexes could be identified by targeting 16S-23S ITS genes [10] , due to the same 16 S-23 SITS sequence of Mycobacterium tuberculosis complex, Mycobacterium marinum and Mycobacterium ulcerans, it is impossible to identify
DNA sequencing, PCR-restriction fragment analysis (PCR-RFLP), but both are not suitable for mixed infections

Method used

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  • Mycobacterium PCR-RLB inspection and its primer and probe
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  • Mycobacterium PCR-RLB inspection and its primer and probe

Examples

Experimental program
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Effect test

Embodiment 1

[0098] Embodiment 1: PCR-RLB detection and identification for SGM

[0099] 1. Material preparation

[0100] 1. Standard strains: including 25 standard strains, 4 Nocardia strains, 5 Rhodococcus strains and 1 Corynebacterium strain.

[0101] 25 standard strains include Mycobacterium tuberculosis H37 Rv, Mycobacterium bovis, BCG, Mycobacterium microti ATCC19422, Mycobacterium africa ATCC25420, Mycobacterium avium ATCC25291, Mycobacterium intracellulare ATCC13950, Mycobacterium scrofula Bacillus ATCC19981, Mycobacterium kansasii ATCC12478, Mycobacterium gastric ATCC15754, Mycobacterium ulcerans ATCC19423, Mycobacterium marinum ATCC927, Mycobacterium simian ATCC25275, Mycobacterium georgia ATCC14470, Mycobacterium secondary ATCC23292, Mycobacterium malmo Bacillus ATCC29571, Mycobacterium asiaticum ATCC25276, Mycobacterium spp ATCC27962, Mycobacterium xenopus ATCC19250, Mycobacterium georgi ATCC15755, Mycobacterium sulja ATCC35799, Mycobacterium haemophilus ATCC29548, Mycobacteriu...

Embodiment 2

[0176] Embodiment two: for the PCR-RLB detection method of RGM

[0177] 1. Material preparation

[0178] 1. Standard strains: 28 RGM standard strains, including: Mycobacterium chelonis ATCC 35752, Mycobacterium abscessus ATCC 19977, Mycobacterium fortuitously ATCC 6841, Mycobacterium parafortuitously ATCC 19686, Mycobacterium smegmatis ATCC 19420, Mycobacterium phlei ATCC 11758 , Mycobacterium flavin ATCC14474, Mycobacterium senegal ATCC 35796, Mycobacterium dist. ATCC 19340, Mycobacterium taereus ATCC 29678, Mycobacterium light yellow ATCC 43909, Mycobacterium aichi ATCC 27280, Mycobacterium duval ATCC 43910, Chita branch Bacillus ATCC 19627, Mycobacterium africanus ATCC33464, Mycobacterium rhodesia ATCC 27024, Mycobacterium suis ATCC 33776, Mycobacterium neogold ATCC 25795, Mycobacterium aureus ATCC 23366, Mycobacterium gardenii ATCC 27726, Mycobacterium thermotolerant ATCC 19527, komossense ATCC 33014, chubuense ATCC 27278, agri ATCC 27406, pulveris ATCC 35154, tokaiense A...

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Abstract

The present invention discloses a PCR-RLB assay process of mycobacteria and the employed primers and probes. A pair of oligonucleotide primers and 35 species or type-specific probes are designed against the characteristics of DNA sequences of different types of mycobacterias. Via PCR-RLB assay process, 17 mycobacterias and Mycobacterium tuberculosis complexes, Mycobacterium Ulcerans / Marinum and 2 subspecies of Mycobacterium Kansasii can be identified for SGM. 14 species or subspecies can be identified for RGM. Compared with traditional identification process of mycobacteria, the present invention is a rapid, simple, practical, specific and sensitive assay method.

Description

[technical field] [0001] The invention relates to a novel molecular biology method for detecting and identifying mycobacteria; in particular, it relates to a PCR-RLB method for detecting mycobacteria and the used primers and probes. [Background technique] [0002] Mycobacterial infections most commonly cause tuberculosis, leprosy, lung infections, and skin infections, among others. Due to the rising incidence of HIV infection and morbidity, opportunistic infections caused by non-tuberculous mycobacteria (Mycobacterium tuberculosis, MOTT) are also on the rise, such as Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium kansasii infection. WHO estimated that there were 12 million tuberculosis patients in the world in 2005 [2] (miller et al, 2002). Because different mycobacterial infections are treated differently clinically, rapid detection and identification of MOTT is particularly important. [0003] Slow growing mycobacterium (slowly growing mycobacterium,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 熊礼宽
Owner 深圳市慢性病防治院
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