A convenient rapid culture assay for the detection of enteroviruses in clinical samples: comparison with conventional cell culture and RT-PCR
- 1 August 2008
- journal article
- Published by Microbiology Society in Journal of Medical Microbiology
- Vol. 57 (8) , 1000-1006
- https://doi.org/10.1099/jmm.0.47799-0
Abstract
A convenient rapid culture assay (RCA) for the detection of enteroviruses was evaluated against RT-PCR using 576 stool and 102 cerebrospinal fluid (CSF) samples. One hundred and ninety stool samples were also tested by conventional cell culture (CCC). The RCA used immunoperoxidase staining of cell culture plates with a blend of monoclonal antibodies (mAbs) against enterovirus VP1 on the second and sixth days after inoculation. This blend was composed of 5D8/1 (Dako) and four ‘in-house’ mAbs. CCC was performed using fluorescence staining with the Enterovirus Screening Set (Chemicon International) for culture confirmation. Detection of enteroviruses by the RCA was more successful in colonic carcinoma (CaCo-2) and rhabdomyosarcoma (RD) cells than in human embryonic lung fibroblasts, HEp2 and A549 cells. The performance of CCC in RD cells was hindered by rapid cell degeneration and non-specific staining of cells during culture confirmation. The sensitivity of the RCA compared to RT-PCR in stool samples was found to be 71 % (115/161) on the second day and 87 % (140/161) on the sixth day. The sensitivity of the RCA in CSF samples was 38 % (22/58) after 2 days and 59 % (34/58) after 6 days. The specificity of the RCA was 100 %. All CCC-positive samples were positive by the RCA. CCC required 3–14 days for virus recovery. In conclusion, the RCA has the same sensitivity as CCC, significantly shortens the time required for the detection of enteroviruses, and prevents pitfalls associated with using RD cells for CCC. For diagnosis of aseptic meningitis in CSF samples, RT-PCR should be performed.Keywords
This publication has 10 references indexed in Scilit:
- Epidemiologic aspects and laboratory features of enterovirus infections in Western Germany, 2000–2005Journal of Medical Virology, 2007
- Comparison of multiple shell vial cell lines for isolation of enteroviruses: A national perspectiveJournal of Clinical Virology, 2006
- Pitfalls in the Diagnosis of Enteroviral Infection in Young ChildrenThe Pediatric Infectious Disease Journal, 2006
- Human rhabdomyosarcoma cells for rapid detection of enteroviruses by shell-vial assayJournal of Medical Microbiology, 2003
- Comparison of mixed cell culture containing genetically engineered BGMK and CaCo-2 cells (Super E-Mix) with RT-PCR and conventional cell culture for the diagnosis of enterovirus meningitisJournal of Clinical Virology, 2002
- A‐549 is a suitable cell line for primary isolation of coxsackie B virusesJournal of Medical Virology, 2001
- Detection of Precytopathic Effect of Enteroviruses in Clinical Specimens by Centrifugation-Enhanced Antigen DetectionJournal of Clinical Microbiology, 2001
- Comparison of rhabdomyosarcoma, buffalo green monkey kidney epithelial, A549 (human lung epithelial) cells and human embryonic lung fibroblasts for isolation of enteroviruses from clinical samplesJournal of Clinical Virology, 1998
- Comparison of a rapid culture method combining an immunoperoxidase test and a group specific anti-VP1 monoclonal antibody with conventional virus isolation techniques for routine detection of enteroviruses in stoolsJournal of Medical Virology, 1998
- Evaluation of an enterovirus group-specific anti-VP1 monoclonal antibody, 5-D8/1, in comparison with neutralization and PCR for rapid identification of enteroviruses in cell cultureJournal of Clinical Microbiology, 1995