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. 1992 Oct;30(10):2529–2534. doi: 10.1128/jcm.30.10.2529-2534.1992

Detection of Norwalk virus in stool by polymerase chain reaction.

X Jiang 1, J Wang 1, D Y Graham 1, M K Estes 1
PMCID: PMC270473  PMID: 1383265

Abstract

A method of reverse transcription (RT) and polymerase chain reaction (PCR) for the detection of Norwalk virus in human stools was developed. A cationic detergent, cetyltrimethylammonium bromide, was found to effectively remove from stool extracts factors that inhibit the RT-PCR assay. The specificities of the tests were shown by hybridization of the amplified DNA with Norwalk virus-specific cDNA probes and a consistent correlation between virus detection in stools and infection of volunteers. RT-PCR detected virus in stool samples diluted 10(-4) and was about 100 times more sensitive than dot blot hybridization. In serial stool samples collected before and at different times after inoculation of 10 volunteers with Norwalk virus, 37 of 55 were positive by RT-PCR, but only 27 were positive by dot blot hybridization (chi 2 = 22.96; P less than 0.001). Further application of this method should allow detection of Norwalk virus in food or environmental samples such as shellfish and shellfish waters.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Appleton H. Small round viruses: classification and role in food-borne infections. Ciba Found Symp. 1987;128:108–125. doi: 10.1002/9780470513460.ch7. [DOI] [PubMed] [Google Scholar]
  2. Gary G. W., Anderson L. J., Keswick B. H., Johnson P. C., DuPont H. L., Stine S. E., Bartlett A. V. Norwalk virus antigen and antibody response in an adult volunteer study. J Clin Microbiol. 1987 Oct;25(10):2001–2003. doi: 10.1128/jcm.25.10.2001-2003.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gouvea V., Allen J. R., Glass R. I., Fang Z. Y., Bremont M., Cohen J., McCrae M. A., Saif L. J., Sinarachatanant P., Caul E. O. Detection of group B and C rotaviruses by polymerase chain reaction. J Clin Microbiol. 1991 Mar;29(3):519–523. doi: 10.1128/jcm.29.3.519-523.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. JONES A. S. USE OF ALKYLTRIMETHYLAMMONIUM BROMIDES FOR THE ISOLATION OF RIBO- AND DESOXYRIBO-NUCLEIC ACIDS. Nature. 1963 Jul 20;199:280–282. doi: 10.1038/199280b0. [DOI] [PubMed] [Google Scholar]
  5. Jiang X., Estes M. K., Metcalf T. G. Detection of hepatitis A virus by hybridization with single-stranded RNA probes. Appl Environ Microbiol. 1987 Oct;53(10):2487–2495. doi: 10.1128/aem.53.10.2487-2495.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kaplan J. E., Feldman R., Campbell D. S., Lookabaugh C., Gary G. W. The frequency of a Norwalk-like pattern of illness in outbreaks of acute gastroenteritis. Am J Public Health. 1982 Dec;72(12):1329–1332. doi: 10.2105/ajph.72.12.1329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  8. Matsui S. M., Kim J. P., Greenberg H. B., Su W., Sun Q., Johnson P. C., DuPont H. L., Oshiro L. S., Reyes G. R. The isolation and characterization of a Norwalk virus-specific cDNA. J Clin Invest. 1991 Apr;87(4):1456–1461. doi: 10.1172/JCI115152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  10. Saksena N. K., Dwyer D., Barré-Sinoussi F. A "sentinel" technique for monitoring viral aerosol contamination. J Infect Dis. 1991 Nov;164(5):1021–1022. doi: 10.1093/infdis/164.5.1021. [DOI] [PubMed] [Google Scholar]
  11. Sarkar G., Sommer S. More light on PCR contamination. Nature. 1990 Sep 27;347(6291):340–341. doi: 10.1038/347340b0. [DOI] [PubMed] [Google Scholar]
  12. Wilde J., Eiden J., Yolken R. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol. 1990 Jun;28(6):1300–1307. doi: 10.1128/jcm.28.6.1300-1307.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Xi J. N., Estes M. K., Metcalf T. G. In situ hybridization for quantitative assay of infectious hepatitis A virus. J Clin Microbiol. 1989 May;27(5):874–879. doi: 10.1128/jcm.27.5.874-879.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Xi J. N., Graham D. Y., Wang K. N., Estes M. K. Norwalk virus genome cloning and characterization. Science. 1990 Dec 14;250(4987):1580–1583. doi: 10.1126/science.2177224. [DOI] [PubMed] [Google Scholar]
  15. Zhou Y. J., Estes M. K., Jiang X., Metcalf T. G. Concentration and detection of hepatitis A virus and rotavirus from shellfish by hybridization tests. Appl Environ Microbiol. 1991 Oct;57(10):2963–2968. doi: 10.1128/aem.57.10.2963-2968.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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