A multiplex loop-mediated isothermal amplification assay for rapid screening of Acinetobacter baumannii and D carbapenemase OXA-23 gene

作者：Yang, Rungong; Zhang, Honghong; Li, Xiaoxia; Ye, Ling; Gong, Meiliang; Yang, Jinghui; Yu, Jihong; Bai, Jie^*

来源：Bioscience Reports, 2018, 38(5): BSR20180425.

DOI：10.1042/BSR20180425

• POLYMERASE-CHAIN-REACTION
• NOSOCOMIAL OUTBREAK
• CLINICAL-OUTCOMES
• ESCHERICHIA-COLI
• RECA GENE
• RESISTANCE
• TIME
• IDENTIFICATION
• DISSEMINATION
• EPIDEMIOLOGY

Background: Acinetobacter baumannii is a health burden responsible for various nosocomial infections, and bacteremia in particular. The resistance of A. baumannii to most antibiotics including carbapenem has increased. OXA-23-producing A. baumannii is the chief source of nosocomial outbreaks with carbapenem-resistant A. baumannii. Successful antibiotic treatment relies on the accurate and rapid identification of infectious agents and drug resistance. Here, we describe a multiplex loop-mediated isothermal amplification (LAMP) assay for simultaneous and homogeneous identification for A. baumannii infection screening and drug-resistance gene detection. Methods: Four primer pairs were designed to amplify fragments of the recA gene of A. baumannii and the oxa-23 gene. The reaction with a 25 mu l of final volume was performed at 63. C for 60 min. For comparative purposes, we used a traditional method of bacterial identification to evaluate assay efficacy. Results: The multiplex LAMP assay enables simultaneous and homogeneous detection of the recA gene of A. baumannii and the oxa-23 gene and requires less than 21 min with no pre-requisite for DNA purification prior to the amplification reaction. The detection is specific to A. baumannii, and the coincidence rate of the multiplex LAMP and the traditional method was 100%. Conclusions: Our data indicate that the multiplex LAMP assay is a rapid, sensitive, simultaneous and homogeneous method for screening of A. baumannii and its drug-resistance gene.

• 出版日期2018-10-31
• 单位中国人民解放军总医院