Triple real-time fluorescent PCR (polymerase chain reaction) detection primer, detection probe, detection kit and detection method for methicillin-resistant staphylococcus aureus

Abstract

The invention discloses a triple real-time fluorescent PCR (polymerase chain reaction) detection primer, a detection probe, a detection kit and a detection method for methicillin-resistant staphylococcus aureus. The detection primer and the detection probe are designed according to nuc, mecA and femA gene sequences respectively; through the detection primer and the detection probe, triple real-time fluorescent PCR detection is performed on a sample according to the detection method; in a primary amplification reaction, the detection on the specificity of the methicillin-resistant staphylococcus aureus in the sample to be detected is completed so as to simply, conveniently, rapidly and accurately judge whether the sample to be detected contains the methicillin-resistant staphylococcus aureus. When the triple real-time fluorescent PCR detection is performed on the sample according to the detection method, the detection is rapid and a process from sample preparation to detection result obtainment can be completed within 8-10 hours; interferences from false positiveness, cross contamination and the like can be avoided; the detection result is reliable, the detection sensitivity is high, and the detection specificity is high; and a beneficial tool is provided for epidemiological survey through the methicillin-resistant staphylococcus aureus.

Description

Technical field: [0001] The invention belongs to the technical field of bioengineering, and in particular relates to triple real-time fluorescent PCR detection primers, probes, detection kits and detection methods for methicillin-resistant Staphylococcus aureus (MRSA). Background technique: [0002] Methicillin-resistant Staphylococcus aureus (MRSA) was first reported by Jevons in the UK in 1961, and now it has become a global pathogenic microorganism. AIDS) is also listed as the three most difficult infectious diseases in the world. At present, commonly used methods for MRSA drug resistance detection include disk diffusion method, broth dilution method, agar screening method, molecular biology diagnosis method, etc. [0003] Methicillin-resistant Staphylococcus aureus (Meth icillin-resistant Staphylococcus aureus, MRSA) and Methicillin-resistant coagulase-negative staphylococci (MRCNS) have become hospital and community infections common pathogenic bacteria, and the incid...

AI Technical Summary

Problems solved by technology

mecA (Methicillin determinant A) is the drug resistance determinant gene of MRSA, which encodes PBP2a, which has a very low affinity with β-lactam antibiotics, and promotes the synthesis of bacterial cell walls and drug resistance. It has been clarified that the mecA gene only exists in staphylococci. Located on a mobile element composed of different integrons, transposons, etc., the mecA gene can be used as a molecular marker of methicillin-resistant staphylococci, but the mecA gene exists in all methicillin-resistant staphylococci and is specific And conservative, so only detection of mecA gene can not be determined as MRSA

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