Rapid detection of enterovirus 71 by real-time TaqMan RT-PCR.
BACKGROUND
Enterovirus 71 (EV71) is the main etiological agent of Hand, Foot and Mouth Disease (HFMD) and has been associated with neurological complications which resulted in fatalities during recent outbreaks in Asia Pacific region.


OBJECTIVE
Develop a real-time TaqMan RT-PCR for rapid detection of EV71.


METHODS
Specific primers and probe were designed based on highly conserved VP1 region of EV71. The sensitivity of the real-time RT-PCR was evaluated with 67 clinical specimens collected from pediatric patients with suspected HFMD.


RESULTS
Our real-time TaqMan RT-PCR showed 100% specificity in detecting EV71 and showed an analytical sensitivity of 5 viral copies. High sensitivity was also achieved in detecting EV71 directly from clinical specimens.


CONCLUSIONS
Real-time TaqMan RT-PCR offers a rapid and sensitive method to detect EV71 from clinical specimens, and will allow quarantine measures to be taken more effectively during outbreaks.
Tan
Eng Lee
EL
Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
Yong
Li Li Gaynor
LL
Quak
Seng Hock
SH
Yeo
Wei Cheng Andrea
WC
Chow
Vincent Tak Kwong
VT
Poh
Chit Laa
CL
D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers; D029821:Enterovirus A, Human / Q000235:genetics / Q000302:isolation & purification*; D006232:Hand, Foot and Mouth Disease / Q000175:diagnosis* / Q000821:virology*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase; D013997:Time Factors
D017931:DNA Primers; D012367:RNA, Viral; D019914:Taq Polymerase