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Journal of Clinical Microbiology, October 2000, p. 3681-3685, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genotyping of Rotaviruses in Environmental Water and Stool Samples in Southern Switzerland by Nucleotide Sequence Analysis of 189 Base Pairs at the 5' End of the VP7 Gene

Franca Baggi* and Raffaele Peduzzi

Istituto Cantonale Batteriosierologico, Lugano, Switzerland

Received 28 December 1999/Returned for modification 31 May 2000/Accepted 27 July 2000

Stool specimens from children (<4 years old) with diarrhea were collected over a 1-year period in Ticino (southern region of Switzerland). During the same period, environmental samples were collected from surface waters in the proximity of major water treatment plants. From treatment plants, samples were collected from the raw sewage and before the release of the treated water. From rivers, samples were collected before and after receiving the treated waters. A single-step reverse transcription (RT)-PCR amplification of the entire VP7 gene from extracted double-stranded RNA was developed. For the water samples, a further nested PCR was necessary to increase sensitivity. All amplified viral products were sequenced, and the sequence profile was compared to that of the VP7 genes of human and animal rotaviruses from GenBank. Rotavirus strains are characterized by outer capsid proteins G (glycoprotein) and P (protease-cleaved protein). Correct G genotyping of viral sequences from stool and water samples was possible by analyzing only 189 bp at the 5' end of the VP7 gene. In the Ticino region, the most predominant G genotype among clinical and water samples was G1. Genotypes G2 and G4 were found only among clinical samples. We also detected rotavirus G1-type sequences in feces from a healthy adult. This finding corroborates the hypothesis that healthy adults act as potential reservoirs for the spread of rotavirus in the environment. In our experiments, this RT-PCR-based method for rotavirus genotyping has proven to be a useful tool for epidemiological investigations.


* Corresponding author. Mailing address: Istituto Cantonale Batteriosierologico, Via Ospedale 6, CH-6904 Lugano, Switzerland. Phone: 41 91 923 25 22. Fax: 41 91 922 09 93. E-mail: franca.baggi{at}ti.ch.


Journal of Clinical Microbiology, October 2000, p. 3681-3685, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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