Quintuple PCR primer for detecting four salmonella serotypes, kit and application thereof

A technology for Salmonella and Salmonella Enteritidis is applied in the field of five-fold PCR primers for detection of four Salmonella serotypes in kits, which can solve the problems of high price, shortage of supply, and cross-agglutination.

Inactive Publication Date: 2019-03-15
舟山出入境检验检疫局综合技术服务中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Serum slide agglutination test, as an early Salmonella typing method, is intuitive and reliable, but it also has shortcomings: (1) Long identification time: It takes 1-2 days to determine the serotype of a Salmonella strain. takes about a week
(2) The strain itself: Some strains mutate from smooth to rough, resulting in self-condensation and cannot be typed
(3) Incorrect results such as cross agglutination
(5) The commercial Salmonella diagnostic serum on the market is expensive, the quality is uneven, the serotypes contained are not complete, and the supply is in short supply
According to the currently retrieved PCR detection methods for Salmonella choleraesuis, none of them can distinguish between Salmonella choleraesuis and Salmonella paratyphi C. additional aids

Method used

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  • Quintuple PCR primer for detecting four salmonella serotypes, kit and application thereof
  • Quintuple PCR primer for detecting four salmonella serotypes, kit and application thereof
  • Quintuple PCR primer for detecting four salmonella serotypes, kit and application thereof

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Experimental program
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Embodiment

[0043] 1 Materials and methods

[0044] 1.1 Pathogens used in the test

[0045] Salmonella typhi (CICC10867), Salmonella paratyphi C (CICC21512), Salmonella typhimurium (CICC21483), Salmonella enteritidis (CICC24119), Salmonella choleraesuis (CICC21493), Vibrio parahaemolyticus (ATCC17802), Staphylococcus aureus (ATCC6538 ), Listeria monocytogenes (ATCC19115), Vibrio vulnificus (VV001), Shigella flexneri (ATCC12022), Escherichia coli (ATCC25922), Vibrio mimicus (VM001), Shigella sonnei (ATCC51592), Staphylococcus epidermidis, Enterococcus (ATCC33186), Citrobacter freundii, Staphylococcus worstii, Klebsiella pneumoniae.

[0046] 1.2 Main instruments and reagents

[0047] Multiplex PCR amplification kit RR062A (Dalian Biotechnology Co., Ltd.), DL1000bp DNAMarker (Dalian Biotechnology Co., Ltd.), agarose (Dalian Biotechnology Co., Ltd.), PCR amplifier, gel imager , Electrophoresis apparatus and supporting electrophoresis tank, high-speed centrifuge.

[0048] 1.3 Primer design...

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Abstract

The invention relates to the technical field of pathogenic bacteria detection and discloses a quintuple PCR primer for detecting four salmonella serotypes, a kit and application thereof. The primer comprises five pairs of specific primers and is characterized in that the five pairs of specific primers are salmonella-F and salmonella-R; salmonella enteritidis-F and salmonella enteritidis-R; salmonella typhimurium-F and salmonella typhimurium-R; salmonella typhi-F and salmonella typhi-R; and salmonella suipestifer-F and salmonella suipestifer-R respectively. The quintuple PCR primer disclosed bythe invention not only can detect whether to be salmonella, but also can simply and accurately identify whether to be salmonella typhimurium, salmonella enteritidis, salmonella typhi and salmonella suipestifer at the same time through PCR detection without the aid of other auxiliary technical means.

Description

technical field [0001] The invention relates to the technical field of detection of pathogenic bacteria, in particular to a five-fold PCR primer for detecting four serotypes of Salmonella, a kit and an application thereof. Background technique [0002] Salmonella (Salmonella) is the most common food-borne pathogen, and the food poisoning cases caused by it often rank first in food poisoning incidents around the world. Salmonella infects approximately 1.4 million people in the United States each year and kills 600. The bacteria are widely distributed and usually live in the intestinal tract of human and animal bodies, and pollute their living environment and food with excrement, thus spreading between human and animal bodies. There are more than 2,600 serotypes that have been isolated worldwide. Among them, the Salmonella serotype strains that cause human food poisoning are mainly concentrated in groups A to D, accounting for 70% of foodborne Salmonella, especially Salmonel...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/10C12R1/42
CPCC12Q1/689C12Q2600/16
Inventor 胡兴娟沈飚徐君辉江玲丽施进王忠发余辉
Owner 舟山出入境检验检疫局综合技术服务中心
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