Article
Optimization of Methods for Detecting Hepatitis A Virus in Food
Registro en:
PAULA, V. S. de; GASPAR, A. M. C.; VILLAR, L. M. Optimization of Methods for Detecting Hepatitis A Virus in Food. Food Environ Virol., v.2, p.47–52, 2010.
1867-0334
1867-0342
Autor
Paula, V. S. de
Gaspar, A. M. C.
Villar, L. M.
Resumen
Hepatitis A virus (HAV) is currently recognized
as an important human food borne pathogen, and it is
one of the most resistant enteric RNA viruses, is highly
infectious, and may lead to widespread outbreaks. The aim
of this study was to optimize the methods to detect HAV
from artificially contaminated food. To this end, strawberry
and lettuce were experimentally contaminated with HAV
suspension containing 6 9 106 copies/ml. After contamination,
HAV persistence and washing procedure were
evaluated at 0, 1, 3, 7, and 9 days of storage. Five elution
buffers (PBS (pH 7.4)/0.1% Tween80; 50 mM glycine/3%
(wt/vol) beef extract (pH 9.5); PBS (pH 7, 4); 25 mM
glycine/0.1 Tween80; and 1 M sodium bicarbonate) were
used to elute the virus, and qualitative and quantitative
PCR were used for HAV detection. HAV was detected by
qualitative and quantitative PCR using any of the five
elution buffers, but PBS was the most effective. Even after
washing, HAV was detected up to 9 days after contamination
by quantitative PCR. Quantitative PCR was more
sensitive than qualitative PCR since samples containing
viral load lower than 1.4 9 103 copies/ml could not be
detected by qualitative PCR. Quantitative PCR can be used
for rapid detection of food borne viruses and will help in
the monitoring and control of food borne disease. 2030-01-01