Nucleic acid reagents, kits, systems and methods for detecting drug resistance of Mycobacterium tuberculosis

A technology for Mycobacterium tuberculosis and drug resistance, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Steps, automatic interpretation of results, simple effect of detection process

Active Publication Date: 2022-02-08
北京卓诚惠生生物科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 reagents, kits, systems and methods for detecting drug resistance of Mycobacterium tuberculosis
  • Nucleic acid reagents, kits, systems and methods for detecting drug resistance of Mycobacterium tuberculosis

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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 62 15 rpoB 511 gt(joe)cagt(joe)cagc...

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Abstract

The present disclosure relates to a nucleic acid reagent, kit, system and method for detecting drug resistance of Mycobacterium tuberculosis, wherein the nucleic acid reagent includes SEQ ID NO.1‑6 which are stored independently of each other or randomly mixed with each other The primers shown and the probe shown in SEQ ID NO.11‑18. The present disclosure establishes nucleic acid reagents, kits, systems and methods for detecting drug resistance of Mycobacterium tuberculosis through the above-mentioned primers and probes, which can realize rapid, comprehensive, sensitive, specific and automatic determination of detection results.

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 Patents(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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