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Journal of Clinical Microbiology, October 2000, p. 3681-3685, Vol. 38, No. 10
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.
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
*
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.
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