Multiple-PCR rapid detection method of salmonella and escherichia coli

Abstract

The invention discloses a multiple-PCR rapid detection method of salmonella and escherichia coli. A primer for performing multiple-PCR rapid detection on salmonella and escherichia coli O78 comprises (1) a nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:5; or (2) a nucleotide sequence having the basic sequence homology with the nucleotide sequence in (1) and having the function as same as that of the nucleotide sequence in (1). According to the detection method, specific genes of salmonella and escherichia coli O78 and a part of sequences of bacteria 16s rDNA are amplified once by using the multiple-PCR technology, and a PCR amplification product is detected by means of agarose gel electrophoresis. According to the method, the problems of long period (4-7 days) of a traditional pathogenic bacteria detection technology, tedious and complicated operations and relatively-low specificity and sensitivity are solved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a multiple PCR rapid detection method and application of Salmonella and Escherichia coli O78. Background technique [0002] Rapid detection and identification of pathogenic bacteria is the prerequisite for timely and effective prevention and control of the spread of pathogenic bacteria. It is of great significance to the healthy and normal development of the livestock and poultry industry, food inspection and quarantine safety, and human health. [0003] At present, pathogenic bacteria in food sanitation microbiology inspection (GB / T 4789.1-2003) still mainly rely on traditional bacterial culture, serology, biochemical identification and other methods. Both are low and cannot meet the needs of rapid detection. The fluorescent PCR method introduced by Applied Biosystems of the United States in 1996 quickly developed into an important means of molecular biology detection d...

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Problems solved by technology

[0003] At present, pathogenic bacteria in food sanitation microbiology inspection (GB / T 4789.1-2003) still mainly rely on traditional bacterial culture, serology, biochemical identification and other methods. are low and cannot meet the needs of rapid detection

The fluorescent PCR method introduced by Applied Biosystems of the United States in 1996 quickly developed into an important means of molecular biology detection due to its advantages of low pollution and high sensitivity. Interference is difficult to predict, and fluorescent probes are expensive, so it is difficult to establish a multiplex fluorescent PCR method, and its practicability is greatly affected

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