title: Rapid and visual detection of human enterovirus coxsackievirus A16 by reverse transcription loop-mediated isothermal amplification combined with lateral flow device.
abstract: In this study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow device (LFD) technology to rapidly detect CVA16 was developed and evaluated. RT-LAMP assay was optimized to amplify VP1 gene of CVA16. Amplified products were analysed by LFD and capillary electrophoresis. The RT-LAMP-LFD assay showed 100% specificity in detecting CVA16, and showed analytical sensitivity of 0·55 TCID50 per reaction mixture. Comparison of the RT-LAMP-LFD assay with real-time RT-PCR developed previously in clinical specimens showed 93·3% agreement. The RT-LAMP-LFD assay is more sensitive in detecting CVA16 RNA. The RT-LAMP-LFD assay presented here might offer a rapid and simple alternative in clinical diagnosis of CVA16.
  
  
  CONCLUSIONS
  Coxsackievirus A16 (CVA16) is one of the major causative agents of hand, foot and mouth disease (HFMD). Rapid and reliable detection and typing of it can limit the spread. We developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow device (LFD) technology to rapidly detect CVA16. The high sensitivity and specificity and its ease of use make this assay ideal for use in resource-limited settings such as primary care facilities and clinical laboratories in developing countries.
authors.affiliation: Department of Neonatology, Nanjing Children's Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: G
authors.initials: G
authors.lastname: Yan
authors.affiliation: Department of Emergency, Nanjing Children's Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: L
authors.initials: L
authors.lastname: Jun
authors.affiliation: Key Laboratories of Enteric Pathogenic Microbiology (Ministry of Health), Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Prevention and Control, Nanjing, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Kangchen
authors.affiliation: Key Laboratories of Enteric Pathogenic Microbiology (Ministry of Health), Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Prevention and Control, Nanjing, China.
authors.forename: G
authors.initials: G
authors.lastname: Yiyue
authors.affiliation: Department of Neonatology, Nanjing Children's Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Neonatology, Nanjing Children's Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Xiaoyu
authors.affiliation: Key Laboratories of Enteric Pathogenic Microbiology (Ministry of Health), Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Prevention and Control, Nanjing, China.
authors.forename: S
authors.initials: S
authors.lastname: Zhiyang
authors.affiliation: Key Laboratories of Enteric Pathogenic Microbiology (Ministry of Health), Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Prevention and Control, Nanjing, China.
authors.forename: C
authors.initials: C
authors.lastname: Lunbiao
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D003384:Coxsackievirus Infections / Q000175:diagnosis* / Q000821:virology; D029821:Enterovirus A, Human / Q000235:genetics* / Q000302:isolation & purification; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral
keywords: RT-PCR; coxsackievirus A16; enterovirus; lateral flow device; reverse transcription loop-mediated isothermal amplification