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Triplex PCR detection kit for Escherichia coli serotypes O26, O45 and O121 and primer set thereof

A serotype detection, Escherichia coli technology, applied in recombinant DNA technology, DNA/RNA fragment, microbial determination/inspection, etc., can solve problems such as laborious and time-consuming, and achieve the effect of low cost, simple operation and great economic benefits

Pending Publication Date: 2016-08-03
SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional hemagglutination test for the O antigen is the main method for detecting serotypes. This method is time-consuming and laborious. PCR detection technology has been developed and applied in the detection process of E. coli serotypes. PCR detection method has not been reported yet, therefore, the researchers of the present invention study a kind of multiplex PCR detection method and be applied to the detection of O26, O45, O121 serotype, this method is time-saving, labor-saving, specificity and sensitivity height, is suitable for promoting application

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  • Triplex PCR detection kit for Escherichia coli serotypes O26, O45 and O121 and primer set thereof
  • Triplex PCR detection kit for Escherichia coli serotypes O26, O45 and O121 and primer set thereof
  • Triplex PCR detection kit for Escherichia coli serotypes O26, O45 and O121 and primer set thereof

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Embodiment 1

[0029] Design of Primer Sets for Detection of Escherichia coli O26, O45, O121 Serotypes

[0030] Three pairs of serotype-specific primers were designed. The 159bp, 241bp and 632bp serotype-specific fragments of O26, O45 and O121 were respectively amplified according to the designed primer set.

[0031] 2. Preparation of Escherichia coli O26, O45, O121 serotype detection kits

[0032] (1) Preparation of E. coli template

[0033] Escherichia coli strains were inoculated into VIB liquid medium and enriched overnight at 37°C. Take 1mL of bacterial liquid each to make a template for later use.

[0034] (2) Preparation of specific primers

[0035]The sizes of target fragments amplified by specific primers for Escherichia coli O26, O45 and O121 detected by this kit are 159bp (O26), 241bp (O45) and 632bp (O121), respectively. The concentration of each primer is 10uM, and the primer dilution is stored at -20°C for future use, avoiding repeated freezing and thawing; it can be artif...

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Abstract

The invention discloses a triplex PCR detection kit for Escherichia coli serotypes O26, O45 and O121 and a primer set thereof. Three pairs of primers of the kit can be effectively amplified in a same reaction system. Experiments show that the detection kit containing the primer set and a detection method of the detection kit have the characteristics of being high in speed, sensitive, specific, low in cost and simple to operate, can be used for preferably covering the shortage in the current detection method for the Escherichia coli serotypes O26, O45 and O121, and satisfying the current detection demands of the Escherichia coli serotypes O26, O45 and O121, is easy to popularize and apply in a large scale, is suitable for the preliminary screening of food-borne pathogenic bacteria, and has broad market prospect and relatively high economic benefit.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a detection kit for Escherichia coli O26, O45 and O121 serotypes and a primer set thereof. Background technique [0002] Escherichia coli (Escherichia coli), commonly known as Escherichia coli, was discovered by Escherich in 1885. For a long period of time, it has been regarded as a component of normal intestinal flora and considered to be a non-pathogenic bacterium. It was not until the middle of the 20th century that some special serotypes of Escherichia coli were recognized to be pathogenic to humans and animals, especially to infants and young animals (poultry), often causing severe diarrhea and sepsis. Shiga toxin-producing Escherichia coli is an important source of foodborne pathogens and seriously endangers public health. O157:H7 can cause clinical symptoms such as diarrhea, hemorrhagic enteritis, and uremia. In recent years, Escherichia coli pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/689C12Q2600/16
Inventor 王金良沈志强陈金龙
Owner SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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