Detection primers for QacB gene in methicillin-resistant staphylococcus aureus (MRSA)

A detection primer and gene detection technology, applied in the field of molecular biology, can solve the problems of cumbersome sequencing technology steps, low sensitivity, and long time consumption, and achieve the effect of preventing bacterial strains from continuing to be popular, high resolution, and high detection rate

Inactive Publication Date: 2012-07-04
上海中优医药高科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 QacB gene in methicillin-resistant staphylococcus aureus (MRSA)
  • Detection primers for QacB gene in methicillin-resistant staphylococcus aureus (MRSA)
  • Detection primers for QacB gene in methicillin-resistant staphylococcus aureus (MRSA)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

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

[0024] QacB gene forward primer: 5'-ggttgtggaagaactttctcctt-3';

[0025] QacB gene reverse primer: 5'-tccaattccggaaggtaaca-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 Yir...

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Abstract

The invention discloses detection primers for a QacB gene in MRSA and a detecting method using the same. The detection primers for the QacB gene in MRSA are characterized by comprising a QacB gene forward primer (5'-ggttgtggaagaactttctcctt-3') and a QacB gene reversed primer (5'-tccaattccggaaggtaaca-3'). The detection method comprises: preparing a clinical sample isolate which is MRSA positive as a detection sample and a MRSA strain without the QacB gene as a positive reference; inactivating the detection sample and the positive reference at high temperature respectively and extracting the DNA of the sample; artificially synthesizing a QacB sequence, and constructing a recombinant plasmid carrying a QacB gene as a positive reference; and performing polymerase chain reaction(PCR) reactions respectively. The primers and the method have high detection rate and high repeatability.

Description

technical field [0001] The invention relates to a detection method for carrying the disinfectant-resistant QacB 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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IPC IPC(8): C12Q1/68C12N15/11C12R1/445
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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