Primer sets for detecting mecA gene of methicillin-resistant staphylococcus aureus

A technology for methicillin-resistant Staphylococcus aureus, which is applied in the field of detection of methicillin-resistant Staphylococcus aureus and rapid quantitative detection kits for detection of methicillin-resistant Staphylococcus aureus, which can solve the problems of expensive equipment and unusability

Inactive Publication Date: 2019-08-09
SHANGHAI UNIV OF MEDICINE & HEALTH SCI +1
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Benefits of technology

This patented technology improves upon Lampase testing by creating better primers that can detect both MRSA (methiciline resistant staphylolysan) strains and MRS Astrapted from Sensitive Enterobacteriaceae bacterial species such as Escherichia coli or Salmonella enteritidis. It also includes an improved process called flushing which helps remove excessive antibiotics during analysis time without affecting other important genes involved in resistance mechanisms like MRSA. Overall this new technology allows researchers to quickly identify drug-resistance mutations associated with these organism's pathogenesis while reducing costs compared to traditional methods used before treatment.

Problems solved by technology

This patented technical problem addressed by the patents relates to developing a quick and efficient way to detect methicilase resistant staphylolysaccharides bacteria (MRSB), specifically LAMBA, at home or abroad without requiring complicated instrumentation or costly procedures.

Method used

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  • Primer sets for detecting mecA gene of methicillin-resistant staphylococcus aureus
  • Primer sets for detecting mecA gene of methicillin-resistant staphylococcus aureus

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Embodiment Construction

[0024] The present invention is further described below.

[0025] Operation steps of the present invention are:

[0026] 1. Extract RNA;

[0027] Using the GeneJET DNA / RNA Purification Kit (Thermo Fisher Scientific, USA), DNA was extracted from the samples according to the instructions.

[0028] 2. Prepare the LAMP detection reaction system, the reaction system includes primers: F3, B3, FIP, BIP;

[0029]

[0030] 3. The extracted DNA template is subjected to a LAMP amplification reaction, and the total reaction volume of the LAMP detection reaction system is 25 μl. Amplify at 63°C for 1 hour, place the reaction tube in a water bath at 80°C for 5 minutes to terminate the reaction, observe the intensity of green fluorescence or quantify the DNA concentration with a real-time turbidimeter.

[0031] 4. Composition of LAMP detection reaction system:

[0032] dNTPs (1.4mM), Tris-HCl (pH = 8.8, 20mM), (NH 4 ) 2 SO 4 (10mM), KCl(10mM), MgSO 4 (8mM), betaine (0.8M), 0.1% Tw...

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Abstract

The invention discloses primer sets for detecting an mecA gene of methicillin-resistant staphylococcus aureus on a severely-infected patient, and further relates to a method for detecting the methicillin-resistant staphylococcus aureus through the primer sets and a quick quantitative detection kit for detecting the methicillin-resistant staphylococcus aureus through the primer sets. According to the primer sets, the method and the kit, the best primer sequence and the best detection method for measuring the methicillin-resistant staphylococcus aureus with the mecA gene are determined, and a quick and convenient LAMP high-throughput detection method for laboratories is built.

Description

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Claims

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Application Information

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Owner SHANGHAI UNIV OF MEDICINE & HEALTH SCI
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