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Multiple-PCR (polymerase chain reaction) detection kit for macrolide drug resistance genes of staphylococcus infecting livestock and use method

A technology of macrolides and detection kits, which is applied in the field of medical molecular biology, can solve the problems of high cost and long time consumption, and achieve the effect of reducing detection cost, good convenience and specificity

Pending Publication Date: 2017-03-08
GUIZHOU UNIV
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Problems solved by technology

[0008] The technical problem to be solved by the present invention is to provide a triple PCR detection kit for rapidly detecting macrolide drug-resistant genes of staphylococci in livestock and poultry, specific primers, and methods of use thereof, which can significantly increase the The detection sensitivity and detection efficiency of cyclic lactone drug resistance can reduce the detection cost, so as to overcome many shortcomings of the existing technology, such as high cost and long time consumption

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  • Multiple-PCR (polymerase chain reaction) detection kit for macrolide drug resistance genes of staphylococcus infecting livestock and use method

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[0037] The multi-PCR detection kit for drug-resistant genes of Staphylococcus macrolides in livestock and poultry contains PCR template preparation reagents and multiple PCR amplification reagents for drug-resistant genes. 2.5mL-5.0mL solution, the multi-PCR amplification reagent of the drug-resistant gene includes 650μL-1300μL of 2×TaqPCR MasterMix, 400μL-800μL of specific primers for the drug-resistant gene, ermA gene, ermB gene, and ermC gene, and sterilized double Distilled water 1mL-2mL, negative control 100μL-200μL, positive control 100μL-200μL.

[0038] The specific upstream and downstream primer sequences of the three drug-resistant genes, ermA gene, ermB gene, and ermC gene, are as follows:

[0039] ermA: For: 5-TACACTTGGCTTAGGATG-3;

[0040] Rev-1: 5-TGACTAAAGAAGCGGTAA-3;

[0041] ermB:For:5-AACGACGAAACTGGCTAA-3;

[0042] Rev-2: 5-CTGTGGTATGGCGGGTAA-3;

[0043] ermC:For:5-TCAAAACATAATATAGATAAA-3;

[0044] Rev-3: 5-GCTAATATTGTTTAAATCGTCA-3.

[0045] The method f...

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Abstract

The invention relates to a triple-PCR (polymerase chain reaction) detection kit for macrolide drug resistance genes of staphylococcus infecting livestock. The kit comprises three pairs of PCR primers for amplifying drug resistance genes of to-be-detected bacteria, a template preparation reagent and a bacterial drug resistance gene multiple-amplification reagent. Compared with a conventional drug sensitive test, the multiple-type method has the advantages that the detection time is shortened from days to one day. Pathogenic bacteria often contain multiple macrocyclic lactonase genes. The technology meets the demands for detecting multiple genes simultaneously, and the detection accuracy and specificity are improved; the technology can be used for detecting three macrolide type drug resistance genes in clinical bacteria isolates.

Description

[0001] field of invention [0002] The invention relates to a multiple PCR detection technology for staphylococcus macrolide drug-resistant genes, which belongs to the field of medical molecular biology, in particular to the multiplex PCR detection technology of staphylococcus macrolide drug-resistant genes in livestock and poultry [0003] PCR detection technology and its application in the detection of staphylococcus drug resistance in livestock and poultry. Background technique [0004] Macrolide antibiotics are a class of antibacterial drugs with the basic chemical structure of a macrolide ring and a similar antibacterial spectrum. They can be further divided into 14, 15, and 16-membered rings according to the number of carbon atoms that make up the macrolide ring. Macrolide antibiotics. Macrolides include first-generation erythromycin, which replaces penicillin, and second-generation clarithromycin, roxithromycin, etc. Macrolide antibiotics are a class of rapid bacterio...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14
CPCC12Q1/689C12Q1/686C12Q2600/16C12Q2537/143C12Q2545/113
Inventor 程振涛马光强周碧君文明王开功岳筠李毅
Owner GUIZHOU UNIV
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