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Multiplex PCR detection method for Salmonella typhimurium and its serovars

A technology for Salmonella typhimurium and a detection method, which is applied in the field of multiplex PCR detection for detecting Salmonella typhimurium and its serovariants, can solve the problems of undiscovered Salmonella typhimurium and its serovariants, etc., and achieves practicality, short detection time, and reduced detection rate. cost effect

Active Publication Date: 2014-10-22
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Find through literature search to prior art, have not yet found the report relevant with multiplex PCR method detection Salmonella typhimurium and its serovars of the present invention

Method used

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  • Multiplex PCR detection method for Salmonella typhimurium and its serovars
  • Multiplex PCR detection method for Salmonella typhimurium and its serovars
  • Multiplex PCR detection method for Salmonella typhimurium and its serovars

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Establishment of multiplex PCR detection method for Salmonella typhimurium and its serovars

[0043] Step 1, design amplification primers according to the conserved sequence in the genomic DNA sequence of Salmonella typhimurium

[0044] Find the 1800-2930 segment of the specific gene STM4495 (the gene sequence number comes from the whole genome sequence of Salmonella typhimurium LT2) from the genome DNA sequence of Salmonella typhimurium, and use it as the detection target gene of Salmonella typhimurium. The gene sequence is as shown in SEQ ID NO: 1 As shown, at the same time, the DNA sequences of the O5 antigen-encoding gene oafA, the H1 antigen-encoding gene fliC and the H2 antigen-encoding gene fljB of Salmonella typhimurium were selected as the detection target genes for its serovars. The gene sequences are as shown in SEQ ID NO: 2, SEQ ID Shown in NO: 3 and SEQ ID NO: 4;

[0045] Input the DNA sequence of the selected gene into the primer design software Primer Pr...

Embodiment 2

[0070] Identification of Suspected Salmonella Strains

[0071] Using the multiplex PCR detection method for Salmonella typhimurium and its serum variants established in Example 1, 150 suspected strains of Salmonella isolated from food samples were detected. The food samples were collected in suburban supermarkets and bazaars in Shanghai. Sample processing and suspected For the isolation of strains, refer to the national standard GB / T4789.4-2010. After the genomic DNA was extracted from 150 isolates, first use the universal primer of 16S rDNA to verify all the extracted DNA templates, and then use the multiplex PCR system in Example 1 to detect and identify the mouse Salmonella typhi, Copenhagen var., 4,[5],12:i:- and 4,12:-:1,2 serotypes.

[0072] At the same time, all the strains were streaked on the BHI plate, cultured at 37°C for 12 hours, and the serotypes of the strains were identified by slide agglutination method (please refer to the product manual and national standard...

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Abstract

The invention discloses a multiplex PCR (Polymerase Chain Reaction) detection method for salmonella typhimurium and serum variants of the salmonella typhimurium and belongs to the technical field of food safety inspection. The detection method comprises the following steps of: (1) designing amplification primers, with specific gene sequences of the salmonella typhimurium as templates; (2) extracting the DNA (Deoxyribonucleic Acid) of a sample, and carrying out amplification by using a PCR method; and (3) detecting amplification products through gel electrophoresis, and judging electrophoresis results. The invention further relates to four pairs of specific identification primers, wherein the sequences of the primers are respectively shown in SEQ ID NO: 5 and SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 7, SEQ ID NO: 9 and SEQ ID NO: 10, and SEQ ID NO: 11 and SEQ ID NO: 12. When the detection method disclosed by the invention is used to detect the salmonella typhimurium and the serum variants of the salmonella typhimurium, the detection time is short, the cost is low, the practicability is better, the detection results are specific, and the result judgment is simple.

Description

technical field [0001] The invention relates to a multiplex PCR detection method and primers in the technical field of food safety inspection, in particular to a multiplex PCR detection method for detecting Salmonella typhimurium and its serum variants. Background technique [0002] Salmonella Typhimurium (Salmonella Typhimurium) is one of the most common types of Salmonella infection in my country, and about 30% of Salmonella infections are related to it. The bacteria can exist in poultry, domestic animals, rodents, wild birds and other animals as well as human intestines, and can be transmitted among each other. Infection mostly occurs in infants and young children, often causing symptoms such as fever, nausea, vomiting and diarrhea. [0003] Hegde, N.V., Cook, M.L., Wolfgang, D.R. etc. published in "Journal of Clinical Microbiology" (Clinical Microbiology Journal) 2005, Volume 43, No. 8, Page 4208-4211 entitled "Dissemination of Salmonella enterica subsp.enterica serova...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/42
Inventor 史贤明刘斌何晓华施春雷
Owner SHANGHAI JIAO TONG UNIV
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