Kit for identifying nucleic acid of mycobacterium pathogeny through multiple PCR (polymerase chain reaction)

A technology of mycobacteria and kits is applied in the field of PCR identification and multiplex PCR identification kits for mycobacterial pathogenic nucleic acids, and achieves the effects of simple operation, large-throughput diagnosis and detection, and reduction of identification time and detection cost.

Inactive Publication Date: 2015-03-04
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are some multiplex PCR methods at home and abroad that can be used to detect and differentiate Mycobacterium tuberculosis and BCG (Lee H.R. et al., 2010), distinguish Mycobacterium tuberculosis complex, avian tuberculosis complex and slow-growing mycobacterium (Meng Xianghong et al., 2007); multiplex PCR method for distinguishing avian Mycobacterium tuberculosis, Mycobacterium tuberculosis, Mycobacterium intracellulare, etc. Mycobacterium, Mycobacterium bovis), BCG, non-pathogenic mycobacteria (Mycobacterium avian tuberculosis, Mycobacterium intracellulare, etc.)

Method used

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  • Kit for identifying nucleic acid of mycobacterium pathogeny through multiple PCR (polymerase chain reaction)
  • Kit for identifying nucleic acid of mycobacterium pathogeny through multiple PCR (polymerase chain reaction)
  • Kit for identifying nucleic acid of mycobacterium pathogeny through multiple PCR (polymerase chain reaction)

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Embodiment 1

[0023] 1. Design of specific primers

[0024] The 16S rRNA gene is a Mycobacterium-specific gene and is highly conserved. The primers were designed according to the conserved region, and the target fragment size was 575bp, which can distinguish Mycobacteria from non-mycobacteria. The primer sequences are:

[0025] 16S rRNA-F: 5'-acggtgggtactaggtgtgggtttc-3';

[0026] 16S rRNA-R: 5'-tctgcgattagcgactaagacttca-3'.

[0027]Mycobacterium tuberculosis BCG lacks the RD1 region gene of the Mycobacterium tuberculosis complex group, and specific primers were designed for the RD1 region gene RV3873, which can distinguish BCG from other members of the Mycobacterium tuberculosis group. The target fragment size of PCR amplification is 255bp, and the primer sequence is:

[0028] Rv3873-F: 5'-gcgttgaccgagatggattat-3';

[0029] Rv3873-R: 5'-gctcatctcacccagttggc-3'.

[0030] Mycobacterium bovis lacks a 12.7kb gene segment relative to Mycobacterium tuberculosis, so a universal primer RV1506...

Embodiment 2

[0055] Embodiment 2 clinical application

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Abstract

The invention provides a kit for identifying nucleic acid of mycobacterium pathogeny through multiple PCR (polymerase chain reaction). The kit comprises primer composition as follows: 5'-acggtgggtactaggtgtgggtttc-3', 5'-tctgcgattagcgactaagacttca-3', 5'-gcgttgaccgagatggattat-3', 5'-gctcatctcacccagttggc-3', 5'-ttccgaatcccttgtga-3', 5'-ggagagcgccgttgta-3' or 5'-agtcgccgtggcttctctttta-3'. The kit has high sensitivity and high specificity, is simple to operate and can realize rapid and large-flux detection for virulent and avirulent mycobacterium.

Description

technical field [0001] The invention belongs to the technical field of biological detection and identification, and relates to a kit for multiplex PCR identification of mycobacterium pathogenic nucleic acid, in particular to the identification of Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium bovis BCG and non-pathogenic nucleic acid. A primer for PCR identification of mycobacteria and a PCR identification method using the primer combination. Background technique [0002] Bovine tuberculosis is a zoonotic chronic wasting infectious disease caused by Mycobacterium bovis and Mycobacterium tuberculosis. Cross-infection between humans and animals has caused the widespread prevalence of the disease, which seriously threatens public health security. The Mycobacterium tuberculosis complex includes Mycobacterium tuberculosis, Mycobacterium bovis, BCG, Mycobacterium microti, Mycobacterium africanum types I and II, Mycobacterium caprae and Mycobacterium canettii. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/32
CPCC12Q1/686C12Q1/689C12Q2537/143
Inventor 张泉谭海明蔡骁垚张文成范乃成王建明
Owner YANGZHOU UNIV
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