Nucleic acid reagent, kit, system and method used for detecting drug resistance of mycobacterium tuberculosis

A technology for Mycobacterium tuberculosis and drug resistance, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of being susceptible to contamination by bacteria and detection costs, and avoid sample extraction Steps, automatic interpretation of results, simple detection process

Active Publication Date: 2019-04-16
北京卓诚惠生生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These three types are automatic non-radioactive detection systems, which are easy to operate and widely used. However, in general, these rapid culture systems still have problems such as being susceptible to contamination by bacteria and high detection costs.

Method used

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  • Nucleic acid reagent, kit, system and method used for detecting drug resistance of mycobacterium tuberculosis
  • Nucleic acid reagent, kit, system and method used for detecting drug resistance of mycobacterium tuberculosis

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Experimental program
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Embodiment

[0076] 1. Primer and probe synthesis

[0077] Sequence synthesis was performed according to the primer sequences shown in Table 1 and the probe sequences shown in Table 2. In the probe, FAM is 6-carboxyfluorescein, JOE is 2,7-dimethyl-4,5-dichloro-6-carboxyfluorescein, TAMRA is 6-carboxytetramethylrhodamine, and CY5 is 5H-indole Jing. The brackets in the probe sequences in Table 2 indicate that the t on the left side of the brackets has a fluorescent label, and the contents in the brackets indicate the choice of fluorescent labels.

[0078] Table 1

[0079]

[0080] Table 2

[0081] detection target

probe sequence

Tm value (℃)

SEQ ID NO

rpoB 533

g t(joe)cccag t(joe)gccgccagtc

60

11

rpoB 531

ag t(fam)gccg t(fam)ctgtcggcgct

69

12

rpoB 526

cggg t(fam)tgttc t(fam)ggtccatgaatt

67

13

rpoB 516

cggg t(fam)tgttc t(fam)ggtccatgaatt

64

14

rpoB 513

ag t(joe)cagc t(joe)gagccaattcatg

...

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Abstract

The invention discloses a nucleic acid reagent, kit, system and method used for detecting drug resistance of mycobacterium tuberculosis. The nucleic acid reagent comprises primers shown in SEQ ID NO.1-6 and probes shown in SEQ ID NO.11-18, wherein the primers and the probes are respectively stored independently or mixed and stored randomly. By utilizing the primers and the probes, the nucleic acidreagent, kit, system and method used for detecting the drug resistance of mycobacterium tuberculosis are provided, and quick, comprehensive, sensitive, specific and automatic judgment for the detecting result can be achieved.

Description

technical field [0001] The present disclosure relates to the field of biotechnology, in particular to a nucleic acid reagent, kit, system and method for detecting drug resistance of Mycobacterium tuberculosis. Background technique [0002] The main pathogen of human tuberculosis is Mycobacterium tuberculosis (MTB), which belongs to the Mycobacterium tuberculosis complex (MTBC) of the genus Mycobacterium. In addition to MTBC, the Mycobacterium genus also includes Mycobacterium leprae and at least 140 species of non-tuberculous mycobacteria (NTM), and at least 40 species of NTM are pulmonary pathogens. In addition to Mycobacterium tuberculosis, MTBC also includes Mycobacterium bovis, Mycobacterium africanum, Mycobacterium microti, M. caprae, M. canetti, and M. pinnipedii, all of which have high genome homology among species and can cause Tuberculosis, but with different phenotypic characteristics and host range. [0003] According to the World Health Organization (WHO) repor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11
CPCC12Q1/686C12Q1/689C12Q2600/166C12Q2600/16C12Q2537/143C12Q2563/107C12Q2545/101
Inventor 王雷马寅佳林笑冬王晓艳张志强
Owner 北京卓诚惠生生物科技股份有限公司
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