Specific detection of enterovirus 71 directly from clinical specimens using real-time RT-PCR hybridization probe assay.
Enterovirus 71 (EV71) is one of the main causative agents of hand, foot and mouth disease (HFMD) in young children. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. Thus, rapid detection of the virus is required to enable measures to be implemented in preventing widespread transmission. Based on primers and probes targeting at the VP1 region, a real-time reverse-transcriptase polymerase chain reaction (RT-PCR) hybridization probe assay was developed for specific detection of EV71 from clinical specimens. Quantitative analysis showed that the assay was able to detect as low as 5 EV71 viral copies and EV71 was detected from 46 of the 55 clinical specimens obtained from pediatric patients suffering from HFMD during the period from 2000 to 2003 in Singapore. This study showed that the single tube real-time RT-PCR assay developed in this study can be applied as a rapid and sensitive method for specific detection of EV71 directly from clinical specimens.
Tan
Eng Lee
EL
Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore.
Chow
Vincent Tak Kwong
VT
Kumarasinghe
Gamini
G
Lin
Raymond Tze Pin
RT
Mackay
Ian M
IM
Sloots
Theo P
TP
Poh
Chit Laa
CL
D002648:Child; D004770:Enterovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006232:Hand, Foot and Mouth Disease / Q000821:virology*; D006801:Humans; D012367:RNA, Viral / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012367:RNA, Viral