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Detection primers for QacC gene in MRSA (methicillin resistant staphylococcus aureus) and detection method thereof

A technology for detection of primers and gene detection, applied in the field of molecular biology, can solve the problems of cumbersome steps, low sensitivity, and low cost of sequencing technology, and achieve the effects of preventing bacterial strains from continuing to be popular, high resolution, and high degree of automation

Inactive Publication Date: 2012-07-18
上海中优医药高科技股份有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

Gel electrophoresis is low cost and easy to operate, but it has low sensitivity and is prone to false negatives; sequencing is the gold standard for detecting gene mutations, but sequencing technology steps are cumbersome, time-consuming, and prone to contamination

Method used

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  • Detection primers for QacC gene in MRSA (methicillin resistant staphylococcus aureus) and detection method thereof
  • Detection primers for QacC gene in MRSA (methicillin resistant staphylococcus aureus) and detection method thereof
  • Detection primers for QacC gene in MRSA (methicillin resistant staphylococcus aureus) and detection method thereof

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Experimental program
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Effect test

Embodiment

[0023] 1. Prepare a set of primers for detecting QacC gene carrying in MRSA by solid-phase phosphoramidite triester method, including:

[0024] QacC gene forward primer: 5'-tgcaacacctacactaaatataactt-3';

[0025] QacC gene reverse primer: 5'-cgaaactacgccgactatga-3'.

[0026] 2. Detection method:

[0027] (1) Take the clinical sample isolates identified as positive for methicillin-resistant Staphylococcus aureus after culture, inactivate them at high temperature, and use a commercial bacterial genomic DNA magnetic bead extraction kit (Shenzhen Yirui Biotechnology Co., Ltd., YP03002) Extract sample DNA, dilute it to 30ul (concentration 10ng / ul), store it at -20°C, and store it for use; standard Staphylococcus aureus strain (preservation unit: China Agricultural Microorganism Culture Collection Management Center, preservation number: Number: 43300), high-temperature inactivation, extract sample DNA with a commercial bacterial genomic DNA magnetic bead extraction kit (Shenzhen ...

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Abstract

The invention discloses a group of detection primers for a QacC gene in MRSA (methicillin resistant staphylococcus aureus) and a detection method using the primers. The detection primers for the QacC gene in the MRSA are characterized by comprising a QacC gene forward primer 5'-tgcaacacctaccactaaatataactt-3' and a QacC gene reverse primer 5'-cgaaactacgccgactatga-3'. The detection method comprises the following steps of: taking clinical isolates which are identified as MRSA positive after culture as detection samples, taking a staphylococcus aureus strain without carrying the QacC gene as a negative control product, respectively performing high-temperature inactivation and extracting DNA (deoxyribonucleic acid) of the samples; artificially synthesizing a QacC sequence and constructing a recombinant plasmid carrying the QacC gene as a positive control product; and respectively performing PCR (polymerase chain reaction). The detection primers disclosed by the invention have the advantages of high detection rate and strong repeatability.

Description

technical field [0001] The invention relates to a detection method for carrying the disinfectant-resistant QacC gene in methicillin-resistant Staphylococcus aureus (MRSA), and belongs to the technical field of molecular biology. Background technique [0002] With the widespread use of antibacterial drugs, the drug resistance of bacteria has become more and more serious, and multi-drug resistant strains have emerged. Studies have found that the active efflux system in multidrug-resistant strains can significantly reduce the concentration of antibacterial drugs in cells, and at the same time, due to the wide range of substrates transported by the active efflux system, the bacteria show a high degree of antibacterial drugs with completely different chemical structures. drug resistance. Further studies have shown that the active efflux of bacteria is the main reason for the multi-drug resistance of bacteria. Studies suggest that the drug resistance of MRSA may be related to th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12N15/11
Inventor 傅咏南张奕王校
Owner 上海中优医药高科技股份有限公司
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