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PCR detection primers and kits for bacterial drug resistance gene ndm-1

A detection kit and drug resistance gene technology, applied in the field of molecular biology, can solve problems such as antibiotic knocking, and achieve the effects of low probability, high specificity and high sensitivity

Inactive Publication Date: 2015-09-16
清远市弘顺农牧有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

Although there are no drug-resistant bacteria carrying the NDM-1 gene in my country's animal husbandry industry, the emergence of NDM-1-resistant bacteria has also sounded the alarm for the abuse of antibiotics in my country's animal husbandry industry

Method used

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  • PCR detection primers and kits for bacterial drug resistance gene ndm-1
  • PCR detection primers and kits for bacterial drug resistance gene ndm-1
  • PCR detection primers and kits for bacterial drug resistance gene ndm-1

Examples

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Embodiment 1

[0025] 1. Specific primer design

[0026] According to the sequence of the bacterial drug resistance gene NDM-1 published by GenBank (accession numbers: AB614355, FN396876, FR820590), select the conserved sequence, design specific primers for the detection of the bacterial drug resistance gene NDM-1, and expect the amplified product fragment The length is 241bp (SEQ ID NO:3), and the primer sequences are:

[0027] Primers:

[0028] NDM-1F: GGTTTGGCGATCTGGTTTTCC (SEQ ID NO: 1);

[0029] NDM-1R: TTGTCCTGATGCGCGTGAGTC (SEQ ID NO: 2).

[0030] 2. Preparation of PCR amplification template and positive quality control

[0031] Artificially synthesize the bacterial drug-resistant gene NDM-1 sequence, and use conventional methods to connect the obtained gene NDM-1 fragment into the T vector, which is the PCR amplification template and positive quality control.

[0032] 3. Gene NDM-1 Optimization of PCR Amplification Reaction Conditions

[0033] PCR Amplification System of...

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Abstract

The invention discloses PCR (polymerase chain reaction) detection primers for a drug resistance gene NDM-1 of bacteria and a detection kit containing the primers. Specific PCR primers of the gene NDM-1 are designed to investigate whether drug resistance bacteria in a farm carries the NDM-1 gene or not, the PCR reaction system and condition are optimized to obtain a PCR detection method with high sensitivity, high specificity and high repeatability for a (the) drug resistance gene NDM-1 of bacteria; and the lowest genome DNA (deoxyribonucleic acid) concentration, which can be detected by the detection method, is 9.18*10<-7>mu g / mu L. The method can be used in early diagnosis and quick screen of drug resistance gene NDM-1 carrying bacteria, epidemiological assessment as means for early diagnosis and quick screen of drug resistance gene NDM-1 carrying bacteria, epidemiological assessment,etc. in livestock and poultry breeding process, and can be applied to monitoring, prevention and control of clinical drug resistance bacteria in current veterinarian.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a detection method for livestock and poultry breeding industry, in particular to a PCR detection primer for drug-resistant gene NDM-1 and its application. Background technique [0002] On August 11, 2010, the British medical community announced that a new type of bacteria had emerged in South Asia, which was resistant to almost all antibiotics and was called a "superbug". Because it first appeared in New Delhi, the capital of India, it is also called New Delhi metallo-β-lactamase (New Delhi metallo-β-lactamase-1, NDM-1). NDM-1 is not a bacterium, but a gene encoding metallo-β-lacta-mase (MBL). Subsequently, Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Acinetobacter baumannii, Citric acid, etc. Bacteria such as bacilli infect people and cause death cases, and the clinical manifestations are super drug resistance. [0003] Bacteria carrying the NDM-1 ge...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2531/113
Inventor 温肖会翟少伦魏文康吕殿红袁洁黄忠贾春玲周秀蓉曾琦雯
Owner 清远市弘顺农牧有限公司
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