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Avian pathogenic Escherichia coli triple PCR (polymerase chain reaction) identification kit and identification method

A technology for Escherichia coli and avian pathogenicity is applied in the field of avian pathogenic Escherichia coli triple PCR identification kits, which can solve the problems of time-consuming, labor-intensive, complicated operation, long time-consuming and the like, and achieves obvious application effects, accurate detection results, Less time consuming effect

Pending Publication Date: 2022-02-01
南京悦联生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a triple PCR identification kit and identification method for avian pathogenic Escherichia coli to solve the problems of time-consuming, cumbersome operation, time-consuming and labor-intensive problems in the current market for pathogenic bacteria detection proposed by the above-mentioned background technology

Method used

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  • Avian pathogenic Escherichia coli triple PCR (polymerase chain reaction) identification kit and identification method
  • Avian pathogenic Escherichia coli triple PCR (polymerase chain reaction) identification kit and identification method
  • Avian pathogenic Escherichia coli triple PCR (polymerase chain reaction) identification kit and identification method

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Embodiment 1

[0029] The present invention provides a technical solution: A triple avian pathogenic E. coli and PCR identification kit identification method for the specific amplification comprises three primers serotypes of avian pathogenic E. coli gene sequence combination, wherein the three serotypes were APEC strain EC164, EC165, EC168;

[0030] Nucleotide sequences O2 capsular gene specific amplification primer pairs and have shown SEQIDNo.1 of SEQIDNo.2;

[0031] Nucleotide sequences O78 capsular gene specific amplification primer pairs and have shown SEQIDNo.3 of SEQIDNo.4;

[0032] Nucleotide sequences O145 capsular gene specific amplification primer pairs and have shown SEQIDNo.5 of SEQIDNo.6;

[0033] PCR kit comprises a 2 × RapidTaqMasterMix (withddH2O);

[0034] PCR kit includes three serotypes positive control sample;

[0035] 3 avian pathogenic E. coli bacteria to inactivate EC164 (O78, positive control), EC165 (O2 positive control), EC168 (O145 positive control);

[0036] A metho...

Embodiment 2

[0043] The present invention provides a technical solution: A triple avian pathogenic E. coli and PCR identification kit identification method for the specific amplification comprises three primers serotypes of avian pathogenic E. coli gene sequence combination, wherein the three serotypes were APEC strain EC164, EC165, EC168;

[0044]Primers for specific amplification of O2 capsular membrane genes have a nucleotide sequence shown in SEQIDNO.1 and SEQIDNO.2, respectively;

[0045] Primers for specific amplification of O78 capsular membrane genes have SEQIDNO.3 and SEQIDNO.4, respectively;

[0046] Primers for specific amplification O145 capsular membrane genes have SEQIDNO.5 and SEQIDNO.6, respectively;

[0047] The PCR kit includes 2 × RAPIDTAQMASTERMIX (WITHDH2O);

[0048] The PCR kit includes a positive control sample of three serotypes;

[0049] Three poultry pathogenic E. coli is an EC164 inactivated bacterial solution (O78 positive control), EC165 (O2 positive control), EC16...

Embodiment 3

[0058] BACKGROUND OF THE INVENTION The present invention provides a technical solution: a triple PCR identification kit and identification method of poultry nicherga and identification methods, including primer for specific amplification of three serotype gene sequences, Among them, the three serotypes of serotypes are EC164, EC165, EC168, respectively;

[0059] Primers for specific amplification of O2 capsular membrane genes have a nucleotide sequence shown in SEQIDNO.1 and SEQIDNO.2, respectively;

[0060] Primers for specific amplification of O78 capsular membrane genes have SEQIDNO.3 and SEQIDNO.4, respectively;

[0061] Primers for specific amplification O145 capsular membrane genes have SEQIDNO.5 and SEQIDNO.6, respectively;

[0062] The PCR kit includes 2 × RAPIDTAQMASTERMIX (WITHDH2O);

[0063] The PCR kit includes a positive control sample of three serotypes;

[0064] Three poultry pathogenic E. coli is an EC164 inactivated bacterial solution (O78 positive control), EC165...

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Abstract

The invention discloses an avian pathogenic Escherichia coli triple PCR (polymerase chain reaction) identification kit and an identification method. The avian pathogenic Escherichia coli triple PCR identification kit comprises a primer pair combination of avian pathogenic Escherichia coli gene sequences of three serotypes for specific amplification, wherein APEC strains of the three serotypes are respectively EC164, EC165 and EC168. The primer pair for specifically amplifying the O2 type capsular gene has nucleotide sequences as shown in SEQ ID No. 1 and SEQ ID No. 2 respectively. The primer pair for specifically amplifying the O78 type capsular gene has nucleotide sequences as shown in SEQ ID No.3 and SEQ ID No.4 respectively. The primer pair for specific amplification of the O145 type capsular gene has nucleotide sequences as shown in SEQ ID No.5 and SEQ ID No.6 respectively. The avian pathogenic Escherichia coli triple PCR identification kit and the identification method have the advantages of good specificity, rapidness, simplicity, convenience and low cost, can rapidly detect predominant epidemic serotypes, grasp epidemiological characteristics and provide a basis for disease prevention and control.

Description

Technical field [0001] The present invention relates to the technical field of detection veterinary infectious diseases, in particular to a triple PCR avian pathogenic Escherichia coli and identification kit identification method. Background technique [0002] Avian pathogenic E. coli ( Avian Pathogenic Escherichia Coli , APEC) is a Gram-negative bacteria, is a kind of intestinal pathogenic E. coli, are widely prevalent in chickens and ducks, geese and other waterfowl, usually associated with sick birds after infection and systemic disease characteristic lesions such as pericarditis, liver Zhou Yan, airbags go far, and even cause acute septicemia and death. Poults susceptible to the disease, usually caused by acute sepsis deaths Typically, the larger day-old chickens have a certain resistance, from an annual layers of septic peritonitis E. coli can cause high mortality and APEC can also penetrate the shell caused the chick embryo infected, resulting in dead embryos, hatching rate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6858C12R1/19
CPCC12Q1/689C12Q1/6858C12Q2600/16C12Q2600/156C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 张炜厉从志王卓昊郑祥宽
Owner 南京悦联生物科技有限公司
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