Bacterium drug-resistant gene detection method, gene chip and kit

A technology for detecting kits and drug-resistant genes, which is applied in the field of biomedicine to achieve significant clinical practical value, fast detection speed, high sensitivity and specificity

Active Publication Date: 2013-03-06
NINGBO NO 2 HOSPITAL +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PCR hybridization analysis is the most common method to detect bacterial drug resistance genes. However, scre

Method used

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  • Bacterium drug-resistant gene detection method, gene chip and kit
  • Bacterium drug-resistant gene detection method, gene chip and kit
  • Bacterium drug-resistant gene detection method, gene chip and kit

Examples

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Embodiment 1

[0028] Example 1 Using the kit of the present invention to detect drug resistance genes of common clinical drug-resistant bacteria

[0029] 1. Detection target and design of primers and probes

[0030] The detection targets of this embodiment include: Staphylococcus aureus and Enterococcus drug resistance genes in Gram-positive cocci, and Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa in Gram-negative bacilli Resistant genes of spp. and Acinetobacter baumannii.

[0031] The drug resistance genes have no bacterial species-specific differences, including TEM, SHV, CTX-M, DHA, CIT, IMP, VIM, KPC, OXA-23, qnrB, qnrS, mecA, aac(6')-Ie- aph(2”)-Ia, aac(6')-Ib, GyrA and ParC, among them, the aac(6')-Ib gene is designed with 3 mutation sites, namely T304C, T304A, G535T; GyrA designed 3 mutations The sites are N87, Y87, and L83; ParC designed 4 mutation sites, namely I80, K84, G84, and V84.

[0032] The sequence length of the forward primer and the reve...

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Abstract

The invention discloses a bacterium drug-resistant gene detection method, a gene chip and a kit. The kit comprises a primer pair for amplifying the bacterium drug-resistant gene to be detected, and detection probes. The gene chip comprises probes for detecting the bacterium drug-resistant gene to be detected. The method for detecting the drug-resistant gene by using the kit and gene chip comprises the following steps: 1) carrying out PCR (polymerase chain reaction) amplification on the drug-resistant gene in the sample DNA (deoxyribonucleic acid) to be detected by using the primers in the kit; 2) carrying out fluorescence labeling reaction on the amplified product and the detection probes in the kit; and 3) hybridizing the fluorescence-labeled amplified product with the gene chip to determine the drug-resistant gene contained in the sample. The detection method disclosed by the invention covers the detection of the most common 16 drug-resistant genes in clinic, and is beneficial to implementing rapid diagnosis of clinical bacterium drug resistance conditions, thereby providing supports for reasonably selecting therapeutic drugs in clinic.

Description

Technical field [0001] The invention relates to the field of biomedicine, in particular to a method for detecting drug resistance genes of common clinical bacteria. The invention also relates to a kit and a gene chip used in the method. Background technique [0002] Statistics from the National Bacterial Resistance Surveillance Network show that among drug-resistant bacteria, Gram-negative bacteria account for about 70%. Among them, Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas aeruginosa, and Bauman bacteria account for about 70%. There are many kinesin bacteria, and the multi-drug resistance is serious; Gram-positive bacteria account for about 30%, of which Staphylococcus accounts for 2 / 3, while Staphylococcus aureus accounts for almost half of the Staphylococcus genus, and Enterococcus spp. About 1 / 4 of Gram-positive bacteria, and mainly Enterococcus faecalis and Enterococcus faecium. [0003] With the increase of infectious diseases, the third and...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/02
Inventor 蔡挺张春秀张顺陈金丝李巧云周佳菁肖华胜
Owner NINGBO NO 2 HOSPITAL
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