Rapid detection of and vancomycin resistance using recombinase polymerase amplification.

Vancomycin-resistant enterococci (VRE), especially , have been a global concern, often causing serious healthcare-associated infections. We established a rapid approach for detecting and vancomycin-resistance genes ( and ) in clinical samples using isothermal recombinase polymerase amplification (RPA) combined with a lateral-flow (LF) strip. Specific RPA primer sets and probes for (to identify the presence of ) and 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 ; 35 type and two type) and stool/rectal-swab samples (63 and 36 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 , , and , and it has the potential to be used as a point-of-care device for VRE therapy and prevention.