Primers and method for detecting merA gene in multidrug-resistant pseudomonas aeruginosa (MDRPA)

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 easy contamination, and achieve the effects 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
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  • Claims
  • 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

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  • Primers and method for detecting merA gene in multidrug-resistant pseudomonas aeruginosa (MDRPA)
  • Primers and method for detecting merA gene in multidrug-resistant pseudomonas aeruginosa (MDRPA)
  • Primers and method for detecting merA gene in multidrug-resistant pseudomonas aeruginosa (MDRPA)

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Embodiment

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

[0024] merA gene forward primer: 5'-cgagcaacccgaacatctac-3';

[0025] merA gene reverse primer: 5'-acttgcggatcggtgaac-3'.

[0026] 2. Detection method:

[0027] (1) Take the clinical sample isolates identified as positive for multidrug-resistant Pseudomonas aeruginosa after culture, inactivate them at high temperature, and use a commercial bacterial genomic DNA magnetic bead extraction kit (Shenzhen Yirui Biotechnology Co., Ltd., YP03002 ) to extract sample DNA, dilute to 30ul (concentration 10ng / ul), store at -20°C, and wait for use; Number: 27853), high-temperature inactivation, extract sample DNA with a commercial bacterial genomic DNA magnetic bead extraction kit (Shenzhen Yirui Biotechnology Co., Ltd., YP03002), dilute to 30ul (concentration 10ng / ul), as a negative control ; artificially synthesized merA sequence (synthesized by Sha...

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Abstract

The invention discloses primers and a method for detecting merA gene in multidrug-resistant pseudomonas aeruginosa (MDRPA). The primers for detecting the merA gene in the MDRPA are characterized by comprising an merA gene forward primer 5'-cgagcaacccgaacatctac-3', an merA gene reverse primer 5'-acttgcggatcggtgaac-3'. The detection method comprises the following steps of: taking a clinical sample isolate which is cultured and identified as MDRPA positive, deactivating at high temperature, and extracting sample DNA serving as a detection sample; taking a pseudomonas aeruginosa strain which does not carry the merA gene, deactivating at high temperature, and extracting sample DNA serving as a negative control; artificially synthesizing an merA sequence, and constructing a recombinant plasmid, which carries the merA gene, serving as a positive control; and respectively performing polymerase chain reaction (PCR). The invention has the advantages of high detection rate and high repeatability.

Description

technical field [0001] The invention relates to a method for detecting the carrying condition of a disinfectant-resistant merA gene in multidrug-resistant Pseudomonas aeruginosa (MDRPA), and belongs to the technical field of molecular biology. Background technique [0002] The application and development of disinfectant antiseptics in daily life have existed for centuries. From the empirical use of copper and silver vessels to store drinking water, the use of vinegar and honey to clean wounds, the use of spices to preserve fish and meat, the use of iodine as a wound disinfectant, the application of chlorine-containing aqueous solutions in obstetrics, the use of phenol as Wound dressings as well as in surgery as antiseptics, and the use of divalent mercury ions as sporicides, the whole spectrum of antiseptics is in constant development. At the beginning of the 20th century, humans further developed biguanides (CRAs) and quaternary ammonium compounds (quateraryammonium compou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/385
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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