Rapid detection of Enterococcus and vancomycin resistance using recombinase polymerase amplification

PeerJ

article

Pimchanok Panpru¹  Arpasiri Srisrattakarn¹  Nuttanun Panthasri²  Patcharaporn Tippayawat¹  Aroonwadee Chanawong¹  Ratree Tavichakorntrakool¹  Jureerut Daduang¹  Lumyai Wonglakorn³  Aroonlug Lulitanond¹ 

[1] Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University;The 8th Regional Medical Science Center;Clinical Microbiology Unit, Srinagarind Hospital, Khon Kaen University

关键词: Enterococcus faecium;    Lateral flow strips;    Recombinase polymerase amplification;    VanA;    VanB;    Vancomycin-resistant enterococci;   

DOI  :  10.7717/peerj.12561

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

Vancomycin-resistant enterococci (VRE), especially Enterococcus faecium, have been a global concern, often causing serious healthcare-associated infections. We established a rapid approach for detecting E. faecium and vancomycin-resistance genes (vanA and vanB) in clinical samples using isothermal recombinase polymerase amplification (RPA) combined with a lateral-flow (LF) strip. Specific RPA primer sets and probes for ddl (to identify the presence of E. faecium) vanA and vanB genes were designed. The RPA reaction was performed under isothermal condition at 37 °C within 20 min and read using the LF strip within a further 5 min. A total of 141 positive blood-cultures and 136 stool/rectal swab samples were tested using RPA-LF method compared to the conventional PCR method. The RPA-LF method exhibited 100% sensitivity in both blood-culture (60 E. faecium; 35 vanA type and two vanB type) and stool/rectal-swab samples (63 E. faecium and 36 vanA type) without cross-reaction (100% specificity). The lower detection limit of the RPA-LF was approximately 10 times better than that of the conventional PCR method. The RPA-LF method is an alternative rapid method with excellent sensitivity and specificity for detecting E. faecium, vanA, and vanB, and it has the potential to be used as a point-of-care device for VRE therapy and prevention.

【 授权许可】

CC BY