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Fluorescent PCR (polymerase chain reaction) primer group and kit for rapidly detecting Salmonella pullorum

A technology of Salmonella pullorum and primer set, applied in the fields of biochemical equipment and methods, microorganisms, recombinant DNA technology, etc., can solve the problems of lack of sensitivity, difficulty in distinguishing similarity, non-specific reaction of glass slide agglutination test, etc. The effect of high reaction efficiency, strong specificity and strong sensitivity

Inactive Publication Date: 2020-02-28
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional isolation and identification methods are complicated and cumbersome. At the same time, Salmonella pullorum, Salmonella typhi and Salmonella enteritidis all belong to the Salmonella D serogroup, and the clinical symptoms are often similar and difficult to distinguish. The slide agglutination test is prone to non-specific reactions. And lack of sensitivity, misjudgment often occurs, so it is urgent to invent a convenient, specific and sensitive Salmonella pullorum rapid detection kit for front-line use

Method used

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  • Fluorescent PCR (polymerase chain reaction) primer group and kit for rapidly detecting Salmonella pullorum
  • Fluorescent PCR (polymerase chain reaction) primer group and kit for rapidly detecting Salmonella pullorum
  • Fluorescent PCR (polymerase chain reaction) primer group and kit for rapidly detecting Salmonella pullorum

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Embodiment 1

[0041] Example 1 Fluorescent PCR primer set for rapid detection of Salmonella pullorum

[0042] The primer set of the present embodiment includes:

[0043] qPCR-F: 5′-CGAACCTGCAACAGCTTTAATAGAAAGC-3′;

[0044] qPCR-R: 5′-CTCGTATTTGGTGGCAGTGATGTTC-3′;

[0045] Probe: 5′-TATT(FAM)AGAG(RNA base)TCTAG-Eclipse-3′;

[0046] The primer set targets the rfbs gene of Salmonella pullorum.

[0047] Among them, Eclipse represents a fluorescent quencher group, and FAM represents a fluorescent label.

Embodiment 2

[0048] Embodiment 2 Fluorescent PCR kit for rapid detection of Salmonella pullorum

[0049] The kit of this embodiment includes a reaction solution, and the reaction solution includes qpcr-F, qpcr-R and Probe;

[0050] qPCR-F: 5′-CGAACCTGCAACAGCTTTAATAGAAAGC-3′;

[0051] qPCR-R: 5′-CTCGTATTTGGTGGCAGTGATGTTC-3′;

[0052] Probe: 5′-TATTAGAGTCTAG-Eclipse-3′;

[0053] The primer set targets the rfbs gene of Salmonella pullorum.

[0054] The reaction solution includes:

[0055] 5U / μl Ex Taq HS 0.125μl;

[0056] 10×Ex Taq Buffer 1.5μl;

[0057] dNTP Mixture 2μl;

[0058] RNase HⅡ enzyme (1:10 dilution) 0.1μl;

[0059] DNA 2.5μl;

[0060] qPCR-F 0.5 μl;

[0061] qPCR-R 0.5 μl;

[0062] Probe 0.25μl;

[0063] h 2 O 17.525 μl.

[0064] The amount of each reagent included in the reaction solution is: 10×Ex Taq Buffer (Mg 2+ plus) 20mM; dNTPMixture 2.5mM; DNA 2.5μl; qpcr-F 10μM; qpcr-R 10μM; Probe 10μM.

[0065] Taking the detection of Salmonella pullorum as an example, ve...

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Abstract

The invention discloses a fluorescent PCR (polymerase chain reaction) primer group for rapidly detecting Salmonella pullorum. The primer group comprises qpcr-F:5'-CGAACCTGCAACAGCTTTAATAGAAAGC-3',qpcr-R:5'-CTCGTATTTGGTGGCAGTGATGTTC-3', Probe:5'-TATTAGAGTCTAG-Eclipse-3', and the primer group aims at an rfbs gene of the Salmonella pullorum. The invention further provides a kit based on the primer group. The primer group and the kit have the advantages of high specificity and sensitivity.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a fluorescent PCR primer set and a kit for rapidly detecting Salmonella pullorum. Background technique [0002] Pullorum disease (PD) is a serious systemic disease caused by Salmonella pullorum, which can be transmitted vertically, mainly infecting chickens, affecting reproductive ability and causing high mortality, and the egg production of hens is infected serious decline, resulting in great economic losses. [0003] The early and rapid detection and identification of Salmonella pullorum is the basis for the prevention, control and purification of the disease. The traditional isolation and identification methods are complicated and cumbersome. At the same time, Salmonella pullorum, Salmonella typhi and Salmonella enteritidis all belong to the Salmonella D serogroup, and the clinical symptoms are often similar and difficult to distinguish. The slide agglutination test is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/10C12N15/11C12R1/42
CPCC12Q1/689C12Q2600/156
Inventor 张建民温俊平廖明任涛徐成刚瞿孝云林琦杰
Owner SOUTH CHINA AGRI UNIV
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