Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii

Shuang Liu; Guangtao Huang; Yali Gong; Xiaojun Jin; Yudan Meng; Yizhi Peng; Junning Zhao; Xiaolu Li; Qin Li

doi:10.1093/burnst/tkaa026

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Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii

更新时间：2022-06-09

- Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii
- Burns & Trauma Vol. 8, Issue 1, (2020)
- 作者机构：
  
  1. Department of Plastic & Burns Surgery
  2. The Affiliated Hospital of Southwest Medical University
  3. Department of Dermatology
  4. Dazhou Central Hospital
  5. Institute of Burn Research
  6. Southwest Hospital
  7. Department of Emergency
  8. Zhejiang University School of Medicine
  9. Sichuan Translational Medicine Center of Chinese Medicine
  10. Sichuan Academy of Chinese Medical Sciences
  11. School of Medicine
  12. Department of Infectious Diseases
- 作者简介：
- 基金信息：
- DOI：10.1093/burnst/tkaa026
  CLC：
- Published：2020，
  
  Published Online：2 September 2020，
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Shuang Liu, Guangtao Huang, Yali Gong, Xiaojun Jin, Yudan Meng, Yizhi Peng, Junning Zhao, Xiaolu Li, Qin Li, Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii, Burns & Trauma, Volume 8, 2020, tkaa026, https://doi.org/10.1093/burnst/tkaa026
DOI：

Shuang Liu, Guangtao Huang, Yali Gong, Xiaojun Jin, Yudan Meng, Yizhi Peng, Junning Zhao, Xiaolu Li, Qin Li, Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii, Burns & Trauma, Volume 8, 2020, tkaa026, https://doi.org/10.1093/burnst/tkaa026 DOI：

摘要

Abstract

BackgroundAcinetobacter baumannii (A. baumannii) is one of the pivotal pathogens responsible for nosocomial infections

especially in patients with low immune response

and infection with carbapenem-resistant A. baumannii has been increasing in recent years. Rapid and accurate detection of carbapenem-resistance genes in A. baumannii could be of immense help to clinical staff.MethodsIn this study

a 15-μL reaction system for recombinase polymerase amplification (RPA) was developed and tested. We collected 30 clinical isolates of A. baumannii from the Burn Institute of Southwest Hospital of Third Military Medical University (Army Medical University) for 6 months and tested antibiotic susceptibility using the VITEK 2 system. A. baumannii was detected based on the blaOXA-51 gene by PCR

qPCR and 15 μL-RPA

respectively. Sensitivity and specificity were evaluated. In addition

PCR and 15 μL-RPA data for detecting the carbapenem-resistance gene blaOXA-23 were comparatively assessed.ResultsThe detection limit of the blaOXA-51 gene by 15 μL RPA was 2.86 CFU/ml

with sensitivity comparable to PCR and qPCR. No positive amplification signals were detected in non-Acinetobacter isolates

indicating high specificity. However

only 18 minutes were needed for the 15 μL RPA assay. Furthermore

an antibiotic susceptibility test showed that up to 90% of A. baumannii strains were resistant to meropenem and imipenem; 15 μL RPA data for detecting blaOXA-23 showed that only 10% (n = 3) of A. baumannii isolates did not show positive amplification signals

and the other 90% of (n = 27) isolates were positive

corroborating PCR results.ConclusionWe demonstrated that the new 15 μL RPA assay for detecting blaOXA-23 in A. baumannii is faster and simpler than qPCR and PCR. It is a promising alternative molecular diagnostic tool for rapid and effective detection of A. baumannii and drug-resistance genes in the field and point-of-care testing.

关键词

Keywords

Acinetobacter baumanniiRecombinase polymerase amplificationRapid detectionCarbapenem-resistance geneblaOXA-51blaOXA-23

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