1887

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.

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2008-08-01
2019-10-14
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References

  1. Bourlet, T., Gharbi, J., Omar, S., Aouni, M. & Pozzetto, B. ( 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 stools. J Med Virol 54, 204–209.[CrossRef]
    [Google Scholar]
  2. Buck, G. E., Wiesemann, M. & Stewart, L. ( 2002; ). 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 meningitis. J Clin Virol 25 (Suppl. 1), S13–S18.
    [Google Scholar]
  3. Chapman, N. M., Tracy, S., Gauntt, C. J. & Fortmueller, U. ( 1990; ). Molecular detection and identification of enteroviruses using enzymatic amplification and nucleic acid hybridization. J Clin Microbiol 28, 843–850.
    [Google Scholar]
  4. Faden, H., Patel, P. H. & Campagna, L. ( 2006; ). Pitfalls in the diagnosis of enteroviral infection in young children. Pediatr Infect Dis J 25, 687–690.[CrossRef]
    [Google Scholar]
  5. King, A. M. Q., Brown, F., Christian, P., Hovi, T., Hyypiä, T., Knowles, N. J., Lemon, S. M., Minor, P. D., Palmenberg, A. C. & other authors ( 2000; ). Family Picornaviridae. In Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses, pp. 657–678. Edited by M. H. V. Van Regenmortel, C. M. Fauquet, D. H. L. Bishop, E. B. Carsten, M. K. Estes, S. M. Lemon, J. Maniloff, M. A. Mayo, D. J. McGeoch, C. R. Pringle & R. B. Wickner. San Diego: Academic Press.
  6. Klespies, S. L., Cebula, D. E., Kelley, C. L., Galehouse, D. & Maurer, C. C. ( 1996; ). Detection of enteroviruses from clinical specimens by spin amplification shell vial culture and monoclonal antibody assay. J Clin Microbiol 34, 1465–1467.
    [Google Scholar]
  7. Kok, T. W., Pryor, T. & Payne, L. ( 1998; ). Comparison of rhabdomyosarcoma, buffalo green monkey kidney epithelial, A549 (human lung epithelial) cells and human embryonic lung fibroblasts for isolation of enteroviruses from clinical samples. J Clin Virol 11, 61–65.[CrossRef]
    [Google Scholar]
  8. Lipson, S. M., David, K., Shaikh, F. & Qian, L. ( 2001; ). Detection of precytopathic effect of enteroviruses in clinical specimens by centrifugation-enhanced antigen detection. J Clin Microbiol 39, 2755–2759.[CrossRef]
    [Google Scholar]
  9. Otero, J. R., Folgueira, L., Trallero, G., Prieto, C., Maldonado, S., Babiano, M. J. & Martinez-Alonso, I. ( 2001; ). A-549 is a suitable cell line for primary isolation of coxsackie B viruses. J Med Virol 65, 534–536.[CrossRef]
    [Google Scholar]
  10. Perez-Ruiz, M., Navarro-Mari, J. M., Palacios del Valle, E. & Rosa-Fraile, M. ( 2003; ). Human rhabdomyosarcoma cells for rapid detection of enteroviruses by shell-vial assay. J Med Microbiol 52, 789–791.[CrossRef]
    [Google Scholar]
  11. Rigonan, A. S., Mann, L. & Chonmaitree, T. ( 1998; ). Use of monoclonal antibodies to identify serotypes of enterovirus isolates. J Clin Microbiol 36, 1877–1881.
    [Google Scholar]
  12. Robert Koch-Institut ( 2006; ). Epidemiologisches Bulletin. Poliomyelitis: Zur Surveillance in Deutschland. Enterovirus-Surveillance als neues Instrument der Überwachung etabliert. Epidemiologisches Bulletin 25, 193–195.
    [Google Scholar]
  13. Roth, B., Enders, M., Arents, A., Pfitzner, A. & Terletskaia-Ladwig, E. ( 2007; ). Epidemiologic aspects and laboratory features of enterovirus infections in Western Germany, 2000–2005. J Med Virol 79, 956–962.[CrossRef]
    [Google Scholar]
  14. Schnurr, D., Yagi, S. & Dondero, M. ( 1996; ). Standardization of enterovirus group reactive monoclonal antibodies. In Abstracts of the Twelfth Annual Clinical Virology Symposium, Clearwater, FL.
  15. She, R. C., Crist, G., Billetdeaux, E., Langer, J. & Petti, C. A. ( 2006; ). Comparison of multiple shell vial cell lines for isolation of enteroviruses: a national perspective. J Clin Virol 37, 151–155.[CrossRef]
    [Google Scholar]
  16. Trabelsi, A., Grattard, F., Nejmeddine, M., Aouni, M., Bourlet, T. & Pozzetto, B. ( 1995; ). 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 culture. J Clin Microbiol 33, 2454–2457.
    [Google Scholar]
  17. Van Doornum, G. J. & De Jong, J. C. ( 1998; ). Rapid shell vial culture technique for detection of enteroviruses and adenoviruses in fecal specimens: comparison with conventional virus isolation method. J Clin Microbiol 36, 2865–2868.
    [Google Scholar]
  18. WHO ( 2004; ). Polio Laboratory Manual. Geneva: World Health Organization.
  19. Wiedbrauk, D. L. & Johnston, S. L. G. (editors) ( 1993; ). Enteroviruses. In Manual of Clinical Virology, pp. 92–97. New York: Raven Press.
  20. Yagi, S., Schnurr, D. & Lin, J. ( 1992; ). Spectrum of monoclonal antibodies to coxsackievirus B-3 includes type- and group-specific antibodies. J Clin Microbiol 30, 2498–2501.
    [Google Scholar]
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