Evaluation of a one-tube RT-PCR system for detection of enteroviruses.
BACKGROUND
A highly sensitive PCR assay for early and rapid detection of enteroviral (EV) RNA in CSF is necessary to investigate the role of EV in acute neurological illnesses.


OBJECTIVE
To evaluate and compare two PCR protocols (Titan one-tube RT-PCR and random primed RT-PCR) for detection of enteroviral RNA in CSF.


METHODS
The PCR protocols were evaluated for lower limit of input detection using log dilutions of five stock EV strains and an isolate of enterovirus-71 in minimum essential medium and three EV stock strains in CSF. The tests were also applied on 77 CSF samples, 46 from patients with suspected acute EV neurological illness and 31 from 'disease controls'.


RESULTS
Even though in the initial virus titration assays there was no statistically significant difference in the limit of input detection by Titan system and the random primed two-step PCR, the latter had a higher positivity rate when used on CSF samples from patients (20/46 vs. 10/46, P<0.01).


CONCLUSIONS
Random primed RT-PCR assay is superior to Titan one-tube RT-PCR for detection of EV RNA in CSF.
Manayani
Darly Joseph
DJ
Department of Clinical Virology, Christian Medical College and Hospital, Vellore 632004, India.
Cherian
Thomas
T
Murali
N
N
Finny
G J
GJ
Green
Jonathan
J
Brown
David
D
Ravi
V
V
Abraham
Mary
M
Sridharan
Gopalan
G
D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000134:cerebrospinal fluid / Q000821:virology*; D006801:Humans; D009422:Nervous System Diseases / Q000134:cerebrospinal fluid / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic