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Primers, kit and method for detecting drug resistance of acinetobacter baumannii

An Acinetobacter baumannii, drug resistance technology, applied in the field of genetic testing, can solve the problems of long time, high requirements for experimental technicians, and high experimental costs, achieve high sensitivity, solve experimental time and finished products, and have good repeatability. Effect

Active Publication Date: 2016-02-24
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Method 1 is the gold standard for bacterial susceptibility determination, which takes a long time and requires high experimental technicians, and is suitable for fine diagnosis of Acinetobacter baumannii drug susceptibility; Method 2 has lower requirements for laboratory personnel, but the cost of the experiment is high

Method used

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  • Primers, kit and method for detecting drug resistance of acinetobacter baumannii
  • Primers, kit and method for detecting drug resistance of acinetobacter baumannii
  • Primers, kit and method for detecting drug resistance of acinetobacter baumannii

Examples

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Embodiment 1

[0046] Whether ComM is inserted directly determines whether the A. baumannii strain has natural competent function.

[0047] The laboratory standard strain of Acinetobacter baumannii does not have the ability to form natural competence. In this example, radioactive isotope-labeled DNA was used to test the DNA uptake ability of different Acinetobacter baumannii strains, so as to evaluate their competence formation ability. The different Acinetobacter baumannii strains used were AcinetobacterbaumanniiATCC17978, AcinetobacterbaumanniiAYE and AcinetobacterbaumanniiA118. Competent cells of different Acinetobacter baumannii strains (10ml; 1-3×10 9 cells / ml) were incubated at 30°C in a mineral medium containing α- 35S-dATP-radiolabeledDNA40ng / ml, take out 0.5ml sample each time, and detect the radioactivity of the cells. The result is as figure 1 shown.

[0048] From figure 1 It can be seen that the laboratory standard strains AcinetobacterbaumanniiATCC17978 and Acinetobacterba...

Embodiment 2

[0051] The recombinant plasmid containing the complete ComM gene was transformed into ATCC17978 bacteria by electroporation to obtain the ATCC17978-pET-comM recombinant strain. In the same manner as in Example 1, the radioactivity of the recombinant strain was detected. The result is as figure 2 shown.

[0052] From figure 2 It can be seen that the ATCC17978-pET-comM recombinant strain obviously obtained the natural competent function.

Embodiment 3

[0054] Acinetobacter baumannii clinically detected are mostly multipledrugresistant (MDR) or extensive drug resistant (extensivedrugresistant, XDR). Aminoglycoside antibiotics; extensively drug-resistant strains refer to those resistant to carbapenem antibiotics in addition to the above three types of antibiotics.

[0055] First of all, it is assumed that this part of multidrug-resistant and extensively drug-resistant Acinetobacter baumannii is due to its complete natural competence, and in the subsequent evolution, it acquires exogenous drug-resistant elements and develops into multidrug-resistant or extensively drug-resistant feature. According to whether Acinetobacter baumannii has natural sensitivity, to a large extent, it is determined by whether the ComM gene is complete or not. The present invention quickly judges whether the ComM gene of Acinetobacter baumannii isolated from clinical trials has been inserted and destroyed. Bacilli have multidrug resistance or extensiv...

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Abstract

The invention relates to the field of gene detection, in particular to primers, a kit and a method for detecting drug resistance of acinetobacter baumannii. The primers for detecting the drug resistance of the acinetobacter baumannii comprises an upstream primer P1 and a downstream primer P2, and the base sequences of P1 and P2 are represented as SEQ ID No.1 and SEQ ID No.2. The kit for detecting the drug resistance of the acinetobacter baumannii contains P1 and P2. According to the method for detecting the drug resistance of the acinetobacter baumannii, whether a to-be-detected sample has multi-drug resistance or extensive drug resistance is judged according to detection of ComM gene integrity of the to-be-detected sample. The upstream primer P1 and a downstream primer P2 are designed according to a ComM gene sequence of the acinetobacter baumannii and have the advantages of high specificity, high sensitivity, good repeatability and the like, and then whether the to-be-detected sample has multi-drug resistance or extensive drug resistance is judged on the basis of the ComM gene integrity.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a primer, a kit and a method for detecting drug resistance of Acinetobacter baumannii. Background technique [0002] Acinetobacter baumannii has a strong ability to acquire drug resistance and clone transmission, and multi-drug resistance, extensive drug resistance, and all-drug resistance Acinetobacter baumannii are prevalent worldwide. According to the data of China CHINET Bacterial Resistance Surveillance Network in 2010, Acinetobacter baumannii accounted for 16.11% of clinically isolated Gram-negative bacteria in 14 teaching hospitals in 10 provinces and cities in my country, second only to Escherichia coli and Klebsiella pneumoniae bacteria, has become one of the most important pathogens of nosocomial infection in my country. In addition, Acinetobacter baumannii has long-term survival ability in vitro, which is easy to cause clone dissemination. Therefore, the prevention and c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/01
CPCC12Q1/689
Inventor 龙泉鑫廖璞张震杜艳芬
Owner CHONGQING MEDICAL UNIVERSITY
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