Real time fluorescent PCR reagent kit for discriminating active Listeria monocytogenes and detection method

A Listeria and real-time fluorescence technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of inability to distinguish the dead and alive state of pathogenic bacteria, incomplete data, etc., and achieves improved detection sensitivity and high Sensitivity, the effect of reducing false positive rate
CN101463387AInactive Publication Date: 2009-06-24ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
Publication Date
2009-06-24
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a real-time fluorescent PCR reagent kit for screening active Listeria monocytogenes. The real-time fluorescent PCR reagent kit mainly comprises specific primers, a fluorescent probe, PCR buffer solution, deoxidant nucleoside triphosphate mixture and DNA polymerase. The sequence of the specific primers and the fluorescent probe is as follows: an upstream primer: 5'- CGGAGGTTCCGCAAAAGATG-3', a downstream primer: 5' -CCTCCAGAGTGATCGATGTT-3' and the fluorescent probe: 5' FAM-AAATCATCGACGGCAACCTC-TAMRA-3', wherein, the FAM is a fluorescent reporter group, and the TAMRA is a fluorescent quenching group. The real-time fluorescent PCR reagent kit has the beneficial effects that the real-time fluorescent PCR reagent kit can effectively screen the life and death state of bacteria and has the high sensitivity and the good specificity of the detection of bacteria, thereby reducing the false positive rate of regular PCR amplification; and the real-time fluorescent PCR reagent kit can realize fast, exact and specific detection towards the active Listeria monocytogenes.
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Description

(1) Technical field

[0001] The invention relates to a real-time fluorescent PCR kit and a detection method for screening active Listeria monocytogenes (Listeria monocytogenes, referred to as Listeria monocytogenes). (2) Background technology

[0002] Listeria belongs to the indeterminate genus of Gram-positive non-bacillus in the microbial classification, and there are 7 species of bacteria in this genus (Listeria monocytogenes L.monocytogenes, Listeria ovis L.iuanuii, Listeria innoculus L.innocua, L.welshimeri, L.seeligeri, L.grayi, L.murray), of which only monocytogenes Listeria is pathogenic to humans. Listeria monocytogenes widely exists in soil, water, plants, human and animal feces, and the symptoms of poisoning are severe. In addition to common gastrointestinal manifestations such as vomiting and diarrhea, it can also erode the human central nervous system (main manifestations meningitis, septicemia, miscarriage and other diseases), causing great damage to the body,...

Claims

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