Abstract
The main objective of this study was to determine the applicability of the polymerase chain reaction (PCR) to detection of hepatitis E virus (HEV) in sewage treatment plants and establishment of the prevalence of hepatitis viral diseases in a population. Epidemics of HEV infection because of inadequate public sanitation have been reported in several developing countries. A procedure for concentration of HEV in sewage samples through adsorption to membrane filters, elution with urea-arginine phosphate buffer, and subsequent reconcentration with magnesium chloride enabled us to concentrate HEV to volumes in the microliter range. HEV-specific cDNA was prepared by reverse transcription of the total RNA extracted from samples. Specific DNA amplification by PCR in combination with slot blot hybridization was used to demonstrate the presence of HEV in sewage samples from the inlets and outlets of three sewage treatment plants. The assay was specific for HEV, and a 240-bp amplified product was visualized by ethidium bromide fluorescence. Sewage samples adjusted to pH 5.0 for adsorption of viruses to membrane filters were PCR positive, while samples adjusted to pH 3.5 were PCR negative.
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- Bradley D. W. Enterically-transmitted non-A, non-B hepatitis. Br Med Bull. 1990 Apr;46(2):442–461. doi: 10.1093/oxfordjournals.bmb.a072409. [DOI] [PubMed] [Google Scholar]
- Khuroo M. S. Hepatitis E: the enterically transmitted non-A, non-B hepatitis. Indian J Gastroenterol. 1991 Jul;10(3):96–100. [PubMed] [Google Scholar]
- Li H. H., Gyllensten U. B., Cui X. F., Saiki R. K., Erlich H. A., Arnheim N. Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature. 1988 Sep 29;335(6189):414–417. doi: 10.1038/335414a0. [DOI] [PubMed] [Google Scholar]
- Ramalingaswami V., Purcell R. H. Waterborne non-A, non-B hepatitis. Lancet. 1988 Mar 12;1(8585):571–573. doi: 10.1016/s0140-6736(88)91362-1. [DOI] [PubMed] [Google Scholar]
- Reyes G. R., Purdy M. A., Kim J. P., Luk K. C., Young L. M., Fry K. E., Bradley D. W. Isolation of a cDNA from the virus responsible for enterically transmitted non-A, non-B hepatitis. Science. 1990 Mar 16;247(4948):1335–1339. doi: 10.1126/science.2107574. [DOI] [PubMed] [Google Scholar]
- Zuckerman A. J. Hepatitis E virus. BMJ. 1990 Jun 9;300(6738):1475–1476. doi: 10.1136/bmj.300.6738.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]