Multiple PCR method for detecting quinolone antibiotics campylobacter jejuni and kit

A technology of Campylobacter jejuni and quinolones, applied in the biological field, can solve the problems of long time consumption and poor sensitivity, and achieve the effects of fast and accurate detection, good sensitivity and simple detection technology

Active Publication Date: 2014-01-22
CHINA AGRI UNIV
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Since the above-mentioned drug sensitivity test is based on culture, it takes a long time, about 2-3 days to get the result, and the sensitivity is poor. Therefore, there is an urgent need for rapid and sensitive experimental methods to detect bacterial drug resistance and guide clinical practice. Treatment options to control the spread and spread of drug-resistant bacteria

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  • Multiple PCR method for detecting quinolone antibiotics campylobacter jejuni and kit
  • Multiple PCR method for detecting quinolone antibiotics campylobacter jejuni and kit
  • Multiple PCR method for detecting quinolone antibiotics campylobacter jejuni and kit

Examples

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Embodiment 1

[0030] Example 1. Integrating mismatch amplification mutation analysis PCR (MAMA-PCR) and multiplex PCR technology for strain identification rapid detection of quinolone-resistant Campylobacter jejuni

[0031] Strains to be tested:

[0032] Salmonella resistant to quinolone antibiotics (No. 395, this laboratory’s own strain collected from China, confirmed by drug susceptibility tests that it is resistant to quinolone antibiotics), Salmonella ATCC13311 (purchased from the American Standard Biological Products Collection), Campylobacter coli (numbered TA-60, this laboratory's own strain collected from China, confirmed by drug susceptibility tests to be quinolone antibiotic-resistant Campylobacter coli), quinolone antibiotic-resistant Campylobacter jejuni DH69, quinolone drugs The sensitive Campylobacter jejuni model strain 11168 (numbered NCTC11168), and the quinolone-sensitive (non-resistant) Campylobacter jejuni standard quality control strain 33560 (numbered ATCC33560).

[0033] Qu...

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Abstract

The invention integrates mispairing amplification mutation analysis PCR (MAMA-PCR) and a multiple PCR technique, and provides a method for rapidly detecting quinolone antibiotics campylobacter jejuni caused by point mutation of nucleotide and a kit. The kit comprises a primer composition with the nucleotide sequence shown by SEQ ID No.1-5 and SEQ ID No.7 in the sequence list and a quality control group primer composition with the nucleotide sequence shown by SEQ ID No.1-6 in the sequence list. By adopting the detection method and the kit, campylobacter jejuni of the quinolone antibiotics can be rapidly identified from suspected bacteria, and complex procedures of the traditional detection method using strain identification and drug sensitive test as the representative are simplified. The method and the kit have favorable specificity and sensitivity, and are very suitable for rapidly detecting suspected bacteria at the frontline to provide accurate medicine instructions.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a molecular biology technique for rapidly detecting quinolone antibiotic-resistant Campylobacter jejuni. Background technique [0002] Campylobacter jejuni is a major food-borne pathogen, and diarrhea caused by Campylobacter jejuni infection ranks first in bacterial diarrhea in many economically developed countries. In developing countries, diarrhea caused by Campylobacter jejuni is second only to E. coli and Shigella. In recent years, as the resistance of Campylobacter jejuni has increased, the degree of contamination in food has increased, and food poisoning incidents caused by it have increased rapidly. The World Health Organization (WTO) has listed the disease as one of the most common infectious diseases. my country's National Foodborne Disease Surveillance Network added monitoring for campylobacter disease in 2003. In nature, Campylobacter jejuni is widely present in th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6858C12Q1/686C12Q2531/113C12Q2537/143Y02A50/30
Inventor 汪洋吴聪明吴辰斌沈建忠
Owner CHINA AGRI UNIV
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