Rapid detection of Enterococcus 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 (RP...

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Bibliographic Details
Published in:PeerJ (San Francisco, CA) CA), 2021-12, Vol.9, p.e12561-e12561, Article e12561
Main Authors: Panpru, Pimchanok, Srisrattakarn, Arpasiri, Panthasri, Nuttanun, Tippayawat, Patcharaporn, Chanawong, Aroonwadee, Tavichakorntrakool, Ratree, Daduang, Jureerut, Wonglakorn, Lumyai, Lulitanond, Aroonlug
Format: Article
Language:English
Subjects:
Antimicrobial agents
Cross-reaction
Drug resistance
Drug resistance in microorganisms
Enterococcus faecium
Genes
Health aspects
Infection
Infectious Diseases
Lateral flow strips
Methods
Microbiology
Nosocomial infections
Pathogens
Polymerase chain reaction
Recombinase
Recombinase polymerase amplification
Rectum
Surveillance
Thermal cycling
VanA
VanB
Vancomycin
Vancomycin-resistant enterococci
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Summary: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.
ISSN:2167-8359
2167-8359
DOI:10.7717/peerj.12561