1887

Abstract

A large proportion of diarrhoeal illnesses in children in developing countries are ascribed to an unknown aetiology because the only available methods, such as microscopy and culture, have low sensitivity. This study was aimed at decreasing the diagnostic gap in diarrhoeal disease by the application of molecular techniques. Faecal samples from 158 children with and 99 children without diarrhoea in a hospital in South India were tested for enteric pathogens using conventional diagnostic methods (culture, microscopy and enzyme immunoassays) and molecular methods (six PCR-based assays). The additional use of molecular techniques increased identification to at least one aetiological agent in 76.5 % of diarrhoeal specimens, compared with 40.5 % using conventional methods. Rotavirus (43.3 %), enteropathogenic (15.8 %), norovirus (15.8 %) and spp. (15.2 %) are currently the most common causes of diarrhoea in hospitalized children in Vellore, in contrast to a study conducted two decades earlier in the same hospital, where bacterial pathogens such as spp., spp. and enterotoxigenic were more prevalent. Molecular techniques significantly increased the detection rates of pathogens in children with diarrhoea, but a more intensive study, testing for a wider range of infectious agents and including more information on non-infectious causes of diarrhoea, is required to close the diagnostic gap in diarrhoeal disease.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.2008/003319-0
2008-11-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/57/11/1364.html?itemId=/content/journal/jmm/10.1099/jmm.0.2008/003319-0&mimeType=html&fmt=ahah

References

  1. Amar C. F., East C. L., Gray J., Iturriza-Gomara M., Maclure E. A., McLauchlin J. 2007; Detection by PCR of eight groups of enteric pathogens in 4,627 faecal samples: re-examination of the English case-control Infectious Intestinal Disease Study (1993–1996). Eur J Clin Microbiol Infect Dis 26:311–323 [CrossRef]
    [Google Scholar]
  2. Berkman D. S., Lescano A. G., Gilman R. H., Lopez S. L., Black M. M. 2002; Effects of stunting, diarrhoeal disease, and parasitic infection during infancy on cognition in late childhood: a follow-up study. Lancet 359:564–571 [CrossRef]
    [Google Scholar]
  3. Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. 1990; Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28:495–503
    [Google Scholar]
  4. Buesa J., Colomina J., Raga J., Villanueva A., Prat J. 1996; Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay. Res Virol 147:353–361 [CrossRef]
    [Google Scholar]
  5. DiStefano D. J., Kraiouchkine N., Mallette L., Maliga M., Kulnis G., Keller P. M., Clark H. F., Shaw A. R. 2005; Novel rotavirus VP7 typing assay using a one-step reverse transcriptase PCR protocol and product sequencing and utility of the assay for epidemiological studies and strain characterization, including serotype subgroup analysis. J Clin Microbiol 43:5876–5880 [CrossRef]
    [Google Scholar]
  6. Green J., Gallimore C. I., Norcott J. P., Lewis D., Brown D. W. 1995; Broadly reactive reverse transcriptase polymerase chain reaction for the diagnosis of SRSV-associated gastroenteritis. J Med Virol 47:392–398 [CrossRef]
    [Google Scholar]
  7. Henriksen S. A., Pohlenz J. F. 1981; Staining of cryptosporidia by a modified Ziehl-Neelsen technique. Acta Vet Scand 22:594–596
    [Google Scholar]
  8. Husain M., Seth P., Broor S. 1995; Detection of group A rotavirus by reverse transcriptase and polymerase chain reaction in feces from children with acute gastroenteritis. Arch Virol 140:1225–1233 [CrossRef]
    [Google Scholar]
  9. Iturriza Gomara M., Wong C., Blome S., Desselberger U., Gray J. 2002; Molecular characterization of VP6 genes of human rotavirus isolates: correlation of genogroups with subgroups and evidence of independent segregation. J Virol 76:6596–6601 [CrossRef]
    [Google Scholar]
  10. Jones G., Schultink W., Babille M. 2006; Child survival in India. Indian J Pediatr 73:479–487 [CrossRef]
    [Google Scholar]
  11. Kang G., Iturriza-Gomara M., Wheeler J. G., Crystal P., Monica B., Ramani S., Primrose B., Moses P. D., Gallimore C. I. other authors 2004; Quantitation of group A rotavirus by real-time reverse-transcription-polymerase chain reaction: correlation with clinical severity in children in South India. J Med Virol 73:118–122 [CrossRef]
    [Google Scholar]
  12. Kosek M., Bern C., Guerrant R. L. 2003; The global burden of diarrhoeal disease, as estimated from studies published between 1992 and 2000. Bull World Health Organ 81:197–204
    [Google Scholar]
  13. Kurugol Z., Geylani S., Karaca Y., Umay F., Erensoy S., Vardar F., Bak M., Yaprak I., Ozkinay F. other authors 2003; Rotavirus gastroenteritis among children under five years of age in Izmir, Turkey. Turk J Pediatr 45:290–294
    [Google Scholar]
  14. Levine M. M. 2006; Enteric infections and the vaccines to counter them: future directions. Vaccine 24:3865–3873 [CrossRef]
    [Google Scholar]
  15. Linton D., Lawson A. J., Owen R. J., Stanley J. 1997; PCR detection, identification to species level, and fingerprinting of Campylobacter jejuni and Campylobacter coli direct from diarrheic samples. J Clin Microbiol 35:2568–2572
    [Google Scholar]
  16. Mathew M., Mathan M. M., Mani K., George R., Jebakumar K., Dharamsi R., Kirubakaran C., Pereira S., Mathan V. I. 1991; The relationship of microbial pathogens to acute infectious diarrhoea of childhood. J Trop Med Hyg 94:253–260
    [Google Scholar]
  17. Monica B., Ramani S., Banerjee I., Primrose B., Iturriza-Gomara M., Gallimore C. I., Brown D. W., Fathima M., Moses P. D. other authors 2007; Human caliciviruses in symptomatic and asymptomatic infections in children in Vellore, South India. J Med Virol 79:544–551 [CrossRef]
    [Google Scholar]
  18. Nataro J. P., Kaper J. B. 1998; Diarrheagenic Escherichia coli . Clin Microbiol Rev 11:142–201
    [Google Scholar]
  19. Noel J. S., Fankhauser R. L., Ando T., Monroe S. S., Glass R. I. 1999; Identification of a distinct common strain of “Norwalk-like viruses” having a global distribution. J Infect Dis 179:1334–1344 [CrossRef]
    [Google Scholar]
  20. Olesen B., Neimann J., Bottiger B., Ethelberg S., Schiellerup P., Jensen C., Helms M., Scheutz F., Olsen K. E. other authors 2005; Etiology of diarrhea in young children in Denmark: a case-control study. J Clin Microbiol 43:3636–3641 [CrossRef]
    [Google Scholar]
  21. O'Ryan M., Prado V., Pickering L. K. 2005; A millennium update on pediatric diarrheal illness in the developing world. Semin Pediatr Infect Dis 16:125–136 [CrossRef]
    [Google Scholar]
  22. Pang X. L., Joensuu J., Hoshino Y., Kapikian A. Z., Vesikari T. 1999; Rotaviruses detected by reverse transcription polymerase chain reaction in acute gastroenteritis during a trial of rhesus-human reassortant rotavirus tetravalent vaccine: implications for vaccine efficacy analysis. J Clin Virol 13:9–16 [CrossRef]
    [Google Scholar]
  23. Ruuska T., Vesikari T. 1990; Rotavirus disease in Finnish children: use of numerical scores for clinical severity of diarrhoeal episodes. Scand J Infect Dis 22:259–267 [CrossRef]
    [Google Scholar]
  24. Vidal M., Kruger E., Duran C., Lagos R., Levine M., Prado V., Toro C., Vidal R. 2005; Single multiplex PCR assay to identify simultaneously the six categories of diarrheagenic Escherichia coli associated with enteric infections. J Clin Microbiol 43:5362–5365 [CrossRef]
    [Google Scholar]
  25. Wani S. A., Nabi A., Fayaz I., Ahmad I., Nishikawa Y., Qureshi K., Khan M. A., Chowdhary J. 2006; Investigation of diarrhoeic faecal samples for enterotoxigenic, Shiga toxin-producing and typical or atypical enteropathogenic Escherichia coli in Kashmir, India. FEMS Microbiol Lett 261:238–244 [CrossRef]
    [Google Scholar]
  26. Xiao L., Escalante L., Yang C., Sulaiman I., Escalante A. A., Montali R. J., Fayer R., Lal A. A. 1999; Phylogenetic analysis of Cryptosporidium parasites based on the small-subunit rRNA gene locus. Appl Environ Microbiol 65:1578–1583
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.2008/003319-0
Loading
/content/journal/jmm/10.1099/jmm.0.2008/003319-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error