A method for rapid detection and identification of Listeria bacteria

Abstract

The invention belongs to the technical field of molecular biological detection, and provides a method for detecting and identifying six pairs of PCR (polymerase chain reaction) primers of six kinds of Listeria bacteria from an environment respectively. The PCR primer pairs are designed according to a specific target gene iap (inhibitor of apoptosis protein) of the Listeria bacteria, the target gene is high in stability and specificity, and the six kinds of Listeria bacteria existing in the environment can be rapidly and accurately identified by virtue of a reasonable PCR system and a gel electrophoresis detection means. The detection method comprises the following steps of (1) extracting a sample DNA (deoxyribonucleic acid), and performing PCR amplification by adopting different primer pairs respectively; (2) judging whether a sample to be detected contains the Listeria bacteria or not according to the length of a PCR amplification product fragment. According to the method, the six kinds of Listeria bacteria in the environment and a food sample can be rapidly, accurately and sensitively detected and identified, so that the efficiency is improved, time is saved, and moreover, the detection cost is lowered.

Description

technical field [0001] The invention relates to a method for rapidly detecting and identifying bacteria of the genus Listeria in the environment, belonging to the technical field of molecular biological detection. Background technique [0002] Listeria spp. Firmicutes, along with Bacillus, Clostridium, Enterococcus, Streptococcus and Staphylococcus; low G+C content (36-42%), aerobic or Facultative anaerobic, asporotic bacteria that produce flagella at 10-25°C and are capable of motility. Currently, there are 6 species of Listeria, namely Listeria monocytogenes (L.monocytogenes), Listeria ovis (L.ivanovii) (L. innocua), L. welshimeri, L. seeligeri, and L. grayi. These six species are widely distributed in the environment, and Listeria monocytogenes and Listeria ovis are pathogenic. Listeria monocytogenes is a zoonotic pathogen that can cause severe septicemia and meningitis, and cause listeriosis with a mortality rate of about 20-30% in humans; food is the main route of hu...

AI Technical Summary

Problems solved by technology

For example, the current method for detecting Listeria in GB4789.30-2010, after two times of enrichment for 42-48 hours after sample treatment, then chromogenic medium screening for 24-48 hours, and purification culture for 24-48 hours Finally, biochemical identification, hemolysis and coordinated hemolysis experiments are carried out. The whole process has cumbersome steps, long time (minimum 120 hours, longest 168 hours), high cost and high work intensity; it is urgent to establish simple operation, fast time and good specificity. Molecular biology methods that do not require special, large, expensive equipment

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