Detection primers and detection method for quaternary ammonium compound agent (QacA) gene carrying condition in methicillin-resistant staphylococcus aureus (MRSA)

A technology for detecting primers and detection methods, which is 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 effects of preventing the continued prevalence of bacterial strains, high detection rate, and avoiding cross-contamination

Active Publication Date: 2012-01-25
利康医药科技江苏有限公司
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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 and detection method for quaternary ammonium compound agent (QacA) gene carrying condition in methicillin-resistant staphylococcus aureus (MRSA)
  • Detection primers and detection method for quaternary ammonium compound agent (QacA) gene carrying condition in methicillin-resistant staphylococcus aureus (MRSA)
  • Detection primers and detection method for quaternary ammonium compound agent (QacA) gene carrying condition in methicillin-resistant staphylococcus aureus (MRSA)

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Embodiment 1

[0025] 1. Prepare a group of detection primers for carrying the QacA gene in MRSA by conventional methods, including:

[0026] QacA gene forward primer: 5′-ggttgtggaagaactttctcctt-3′;

[0027] QacA gene reverse primer: 5'-ccaattccggaaggtaacac-3'.

[0028] 2. A detection method for carrying the QacA gene in MRSA, the specific steps are:

[0029] (1) Take the clinical sample isolates identified as positive for methicillin-resistant Staphylococcus aureus after culture, inactivate at high temperature, and use a commercial bacterial genomic DNA magnetic bead extraction kit (Shenzhen Yirui Biotechnology Co., Ltd., YP03002) Extract sample DNA, dilute to 30ul (concentration: 10ng / ul), store at -20°C until use; inactivate the standard Staphylococcus aureus strain (preservation number: ATCC Number: 43300) at high temperature, and use commercial bacterial genome DNA magnetic bead extraction kit (Shenzhen Yirui Biotechnology Co., Ltd., YP03002) was used to extract sample DNA and dilute ...

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Abstract

The invention provides detection primers and a detection method for quaternary ammonium compound agent (QacA) gene carrying condition in methicillin-resistant staphylococcus aureus (MRSA). The detection primers for the QacA gene carrying condition in the MRSA are characterized by comprising a QacA gene forward primer: 5'-ggttgtggaagaactttctcctt-3'; and a QacA gene reverse primer: 5'-ccaattccggaaggtaacac-3'. The detection method comprises the following steps of: taking a clinical sample separation strain which is positive in the MRSA by identification after culture, performing high-temperatureinactivation, and extracting sample DNA serving as a detection sample; taking a staphylococcus aureus strain without carrying QacA genes, performing high-temperature inactivation, and extracting sample DNA serving as a negative control; artificially synthesizing a QacA sequence, and constructing recombinant plasmids carrying QacA genes to obtain a positive control; and performing polymerase chainreaction (PCR). The detection primers and the detection method have the advantages of high detection rate and strong repeatability.

Description

technical field [0001] The invention relates to a detection primer and a detection method for carrying the disinfectant-resistant QacA gene in methicillin-resistant Staphylococcus aureus (MRSA), belonging to the technical field of molecular biology. Background technique [0002] The qac transport system of Staphylococcus aureus belongs to the major facilitator superfamily (MFS), composed of QacA and qacR genes, encoded by plasmids, QacA is a multidrug transporter, because it mediates the transfer of lipophilic disinfectant quarter It is named after the efflux of amine compounds (quaternary ammonium compounds, Qac); QacR is a multidrug binding protein of 188 amino acids, which belongs to the TerR class of transcriptional repressor proteins. When there is no induction substrate, QacR binds to the pseudo-palindromic sequence (IR1) downstream of the QacA promoter and inhibits the transcription of QacA. Because the pseudo-palindromic sequence (IR1) overlaps with the transcripti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12R1/445
Inventor 张奕王校傅咏南
Owner 利康医药科技江苏有限公司
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