LAMP method for detecting enterobacteriaceae food-borne pathogenic bacteria, nucleic acid and primer pairs

Abstract

The invention relates to an LAMP method for detecting enterobacteriaceae food-borne pathogenic bacteria, nucleic acid and primer pairs and belongs to the technical field of food safety detection.The LAMP method includes the following steps that a common gene sequence of the enterobacteriaceae pathogenic bacteria is screened out through bioinformatics and comparative genomics, and the specific amplification primer pairs are designed according to the sequence; an LAMP detection system is established by optimizing reaction conditions.The invention further relates to nucleic acid with the base sequence shown in SEQ ID NO:1 and a group (three pairs) of primers.The base sequences of the primers are shown in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7.Compared with the prior art, the detection method is used for detecting enterobacteriaceae, detection time is short, cost is low, higher practicality is achieved, the detection result is specific, and result judgment is simple.

Description

technical field [0001] The invention relates to the technical field of food safety detection, in particular to a LAMP method for detecting enterobacteriaceae food-borne pathogenic bacteria, a nucleic acid and a primer pair. Background technique [0002] Enterobacteriaceae (Enterobacteriaceae) is defined in "Bergey's Bacterial System Identification Manual" as a large group of Gram-negative non-bacillus bacteria with similar biological characteristics that widely exist in the intestinal tract of humans and animals and in nature. Such bacteria can form obvious colonies on VRBGA agar at 30°C±1°C or 37°C±1°C and can ferment glucose, oxidase-negative bacteria. "Bergey's Handbook of Systematic Bacteriology (2004 Edition)" divides Enterobacteriaceae bacteria into 44 genera and more than 170 species. Enterobacteriaceae now includes bacteria in the coliform group, as well as the pathogenic genera Salmonella, Shigella, Klebsiella, Proteus, and Enterobacter in addition to the coliform ...

AI Technical Summary

Problems solved by technology

The method of ISO21528-1:2004 and SN0738-1997 to detect Enterobacteriaceae is to count MPN, and GB / T4789.31-2003 formulated the phage detection method of Enterobacteriaceae, but in view of the fact that the industry standard needs to count the phages on the VRBGA plate Colonies undergo glucose fermentation and oxidase confirmation tests, and need to prepare medium and prepare sterilized plates. After plate culture, manual counting is required, which may cause errors such as missed detection.

It is relatively difficult for some laboratories to prepare phages in the national standard

The NMKL method only confirms Enterobacteriaceae with negative oxidase. This method can misjudge Acinetobacter wolfii, which does not ferment glucose but can grow with peptone and is also negative for oxidase, as Enterobacteriaceae, resulting in false positives. As a result, the counting result is high; the ISO method is positive for Enterobacteriaceae by oxidase negative and yellow glucose agar, this rule is easy to turn the surface of glucose agar into blue Enterobacter cloacae, Enterobacter aerogenes and Serratia Bacteria missed detection, false negatives occur, resulting in low count results, and these methods have a long detection cycle

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