'RETCIF': a rapid, sensitive method for detection of viruses, applicable for large numbers of clinical samples.
Rapid detection of viruses in clinical samples is important for continuing appropriate antiviral treatment and discontinuing unnecessary antibacterial treatment, as well as for excluding viral pathogens. Yet detection of viral agents may require numerous susceptible cell lines. Even with the shell vial culture method, it is cumbersome for handling large volumes of specimens. A procedure has been developed, which is time and cost-saving and uses specific cell lines in a 96-well microtitre plate and monoclonal antibodies (RETCIF-rapid enhanced tissue culture immunofluorescence). Each clinical sample was inoculated into 12 different wells with five different cell lines. Enhancement was achieved by sonication, centrifugation and hormonal supplementation to the medium used. Cytomegalovirus (CMV), herpes simplex virus (HSV) and respiratory viruses were detected by monoclonal antibodies on day 2, whilst varicella zoster virus (VZV) and enteroviruses were detected on days 5 and 7, respectively. During July-December 1998, 3298 patient specimens were compared by RETCIF and a modified shell vial method. Either or both methods isolated 779 viruses (24% positivity rate), whilst both methods detected 621. Of the 779 viruses, 87% (679) were isolated by the shell vial method in an average time of 4.9 days. For RETCIF the respective rate was 92.5% (721), in an average time of 3.0 days. The RETCIF method is a time-saving procedure, with higher isolation rates than the shell vial method.
Alexander
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R
Department of Microbiology & Infectious Diseases, Women's & Children's Health Care Network, Royal Children's Hospital, Flemington Road, Parkville, 3052, Vic., Melbourne, Australia. alexandr@cryptic.rch.unimelb.edu.au
Lamb
D
D
White
D
D
Wentzel
T
T
Politis
S
S
Rijnsburger
J
J
van Ruyven
D
D
Kelly
N
N
Garland
S M
SM
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002478:Cells, Cultured; D002498:Centrifugation; D005455:Fluorescent Antibody Technique / Q000191:economics / Q000379:methods*; D006801:Humans; D012680:Sensitivity and Specificity; D013010:Sonication; D013997:Time Factors; D014780:Viruses / Q000276:immunology* / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral