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Primer, probe, method and kit for detecting carbapenem antibiotic resisting gene of baumanii

A technology of Acinetobacter baumannii and carbapenems, applied in the field of molecular biology, can solve the problems of low sensitivity, cumbersome operation, time-consuming and laborious, etc., and achieve the effects of high accuracy, high sensitivity and strong specificity

Inactive Publication Date: 2016-12-07
宁波基内生物技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a primer, a probe, a method and a kit for detecting carbapenem-resistant genes of Acinetobacter baumannii, which solve the problem of identifying Acinetobacter baumannii by bacterial culture identification and drug sensitivity test methods in the prior art and carbapenem-resistant Acinetobacter baumannii CRAB, not only the operation is cumbersome, time-consuming and laborious, the sensitivity is low, the repeatability is poor, and the technical problems of the molecular biological mechanism caused by the resistance cannot be understood

Method used

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  • Primer, probe, method and kit for detecting carbapenem antibiotic resisting gene of baumanii
  • Primer, probe, method and kit for detecting carbapenem antibiotic resisting gene of baumanii
  • Primer, probe, method and kit for detecting carbapenem antibiotic resisting gene of baumanii

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Embodiment 1

[0024] The embodiment of the present invention provides a primer and a probe for detecting the carbapenem-resistant gene of Acinetobacter baumannii, and the nucleotide sequence of the primer for detecting the oxacillinase OXA51 gene of Acinetobacter baumannii is as shown in SEQ ID NO: 2 As shown in ~3, the nucleotide sequence of the Taqman probe for detecting the oxacillinase OXA51 gene of Acinetobacter baumannii is shown in SEQ ID NO: 4; the primer core for detecting the oxacillinase OXA23 gene resistant to carbapenem antibiotics The nucleotide sequence is shown in SEQ ID NO: 6-7, and the nucleotide sequence of the Taqman probe for detecting the carbapenem-resistant antibiotic oxacillinase OXA23 gene is shown in SEQ ID NO: 8;

[0025] Wherein, the sequence of the specific primer designed for the oxacillinase OXA51 gene of Acinetobacter baumannii is as follows:

[0026] Upstream primer P-OXA51-F: 5'-TTTATCAAGATTTAGCTCGT-3'

[0027] Downstream primer P-OXA51-R: 5'-AATTATCGACTT...

Embodiment 2

[0037] In the embodiment of the present invention, a method for detecting the carbapenem-resistant gene of Acinetobacter baumannii is provided, including:

[0038] Step 101, sample nucleic acid preparation to obtain a nucleic acid template;

[0039] Step 102, designing specific primers and fluorescently labeled probes for the oxacillinase OXA51 gene of Acinetobacter baumannii and the carbapenem-resistant oxacillinase OXA23 gene, respectively;

[0040]Wherein, the nucleotide sequence of the primer for detecting Acinetobacter baumannii is shown in SEQ ID NO: 2-3, and the nucleotide sequence of the Taqman probe for detecting Acinetobacter baumannii is shown in SEQ ID NO: 4; The nucleotide sequence of the primer for the gene antibiotic gene is shown in SEQ ID NO: 6-7, and the nucleotide sequence of the Taqman probe for detecting the gene resistant to carbapenem antibiotic is shown in SEQ ID NO: 8.

[0041] Step 103, configuring an enzyme reagent, wherein the enzyme is composed of...

Embodiment 3

[0050] The embodiment of the present invention also provides a kit for detecting the carbapenem-resistant gene of Acinetobacter baumannii, including a nucleic acid extraction solution, a first primer-probe mixture, a second primer-probe mixture, and a PCR Reaction enzyme system, negative control substance, positive control substance and the packing box that separate and pack these reagent bottles or tubes collectively, wherein, the first primer probe mixed solution is made of deoxyribonucleoside triphosphate dN(U)TP, Bowman's Composed of the upstream and downstream primers of the oxacillinase OXA51 gene of the Actinobacillus oxacillinase and a fluorescently labeled probe, the second primer-probe mixture consists of deoxyribonucleoside triphosphate dN(U)TP, carbapenem-resistant antibiotic oxacillin Composed of upstream and downstream primers and a fluorescent labeling probe for the gene of cilinase OXA23;

[0051] Wherein, the sequence of the upstream and downstream primers spe...

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Abstract

The invention relates to the technical field of molecular biology, and discloses a primer, probe, method and kit for detecting a carbapenem antibiotic resisting gene of baumanii. A Taqman probe real-time fluorescence PCR method is adopted, and the primer and the fluorescence labeling probe are designed aiming at OXA51 and OXA23 nucleic acid conserved domains respectively. After the primer and the probe are prepared into PCR detecting mixed liquid, enzymes and sample nucleic acid are added, an FAM channel on a PCR instrument is selected for amplification, and the target gene is detected through changes of fluorescence signals. The primer, probe, method and kit have the advantages of being high in accuracy, specificity and sensitivity, and baumanii and CRAB in a clinical sample can be fast and accurately detected.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a primer, a probe, a method and a kit for detecting the carbapenem-resistant gene of Acinetobacter baumannii. Background technique [0002] Acinetobacter baumannii is widely distributed in the hospital environment and is currently an important pathogen causing nosocomial infections. It mainly causes respiratory tract infections, and can also cause sepsis, urinary system infections, and secondary meningitis. The most common site of nosocomial infection is the lung, and it is an important pathogenic bacterium of hospital-acquired pneumonia, especially ventilator-associated pneumonia. Acinetobacter baumannii has the ability to survive in vitro for a long time, and it is easy to cause clone dissemination. [0003] Risk factors for Acinetobacter baumannii infection include: prolonged hospitalization, admission to intensive care unit, receiving mechanical ventilation, invasi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/689C12Q1/686C12Q2561/101C12Q2521/531C12Q2563/107
Inventor 高华山倪剑锋史俊颖王伟建翁毅
Owner 宁波基内生物技术有限公司
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