scholarly journals ENTERIC VIRUSES AS CONTAMINANTS AND BIOINDICATORS IN ENVIRONMENTAL SAMPLES: A REVIEW

2015 ◽  
Vol 20 (2) ◽  
Author(s):  
Gislaine Fongaro ◽  
Hugo Deleon Silva ◽  
El Mahdy Mohamed El Mahdy ◽  
Maria Elisa Magri ◽  
Camila Daminelli Schissi ◽  
...  
1998 ◽  
Vol 38 (12) ◽  
pp. 83-86 ◽  
Author(s):  
K. J. Schwab ◽  
F. H. Neill ◽  
M. K. Estes ◽  
R. L. Atmar

Current methods for the detection of nucleic acid from enteric viruses in environmental samples usually involve extensive concentration and purification of target viruses followed by RT-PCR amplification using two enzymes, reverse transcriptase and Taq polymerase. We have developed a modified method that improves RT-PCR assays by: (i) the use of an RT-PCR internal standard control RNA to identify potential false negative results caused by inhibition of RT-PCR enzymes; (ii) the use of rTth (Perkin-Elmer, Foster City, CA), a heat-stable enzyme that functions as both a reverse transcriptase and DNA polymerase in a single-tube, single-buffer, elevated-temperature reaction; and (iii) the use of thermolabile uracil N-glycosylase (HK-UNG) (Epicentre Technologies, Madison, WI) to prevent PCR product carryover contamination. The new method was compared to the traditional two-enzyme, RT-PCR method for detection of Norwalk virus (NV) and hepatitis A virus (HAV) in buffer, stool, clam and oyster samples. The new method was at least as sensitive in NV and HAV detection compared to the traditional two-enzyme method. The internal standard control successfully detected inhibitors to RT-PCR amplification. NV and HAV PCR products generated with dUTP replacing dTTP during amplification were seeded into subsequent samples to test the prevention of PCR product carryover contamination by HK-UNG. The new method successfully eliminated PCR product carryover contamination in contrast to the traditional two-enzyme method. These improvements to viral nucleic acid detection have the potential to improve sensitivity, specificity and confidence in RT-PCR results.


2010 ◽  
Vol 14 ◽  
pp. e63-e64
Author(s):  
V. Cannella ◽  
G. Purpari ◽  
A. Ferrari ◽  
A. Migliazzo ◽  
P. Di Marco ◽  
...  

2010 ◽  
Vol 109 (3) ◽  
pp. 1093-1104 ◽  
Author(s):  
K. Sdiri-Loulizi ◽  
M. Hassine ◽  
Z. Aouni ◽  
H. Gharbi-Khelifi ◽  
S. Chouchane ◽  
...  

1999 ◽  
Vol 62 (6) ◽  
pp. 691-697 ◽  
Author(s):  
GARY P. RICHARDS

Enteric viruses, including hepatitis A, Norwalk, and Snow Mountain viruses, Hawaii agent, and rotaviruses have been associated with outbreaks of foodborne illness. Classical culturing procedures are available for poliovirus; however, hepatitis A, Norwalk, and many of the other viruses and agents cannot be propagated in cell culture, therefore, molecular biological tools have emerged as a possible means to detect enteric viruses in foods and environmental samples. There are limitations however in the application of polymerase chain reaction and reverse transcription polymerase chain reaction that restrict their usefulness for measuring the virological safety of foods. The most serious limitation is that molecular techniques fail to discriminate between viable and inactivated viruses even though inactivated viruses pose no threat to the consumer and may be present at levels substantially higher than the virulent forms. Other disadvantages include a lack of assay sensitivity and specificity, high assay costs, and a level of technical expertise not available in most food-testing laboratories. Overall, scientific advances in the development of molecular biological tools have outpaced the demonstration of their validity in assessing the virological safety of foods.


2008 ◽  
Vol 75 (2) ◽  
pp. 297-307 ◽  
Author(s):  
Roberto A. Rodríguez ◽  
Ian L. Pepper ◽  
Charles P. Gerba

2010 ◽  
Vol 61 (2) ◽  
pp. 317-322 ◽  
Author(s):  
H. B. Bennett ◽  
H. D. O'Dell ◽  
G. Norton ◽  
G. Shin ◽  
F.-C. Hsu ◽  
...  

Human enteric viruses are important agents of waterborne illness. They are diffusely distributed in environmental waters, necessitating concentration of tens to hundreds of litres for effective detection. This study evaluates the novel ViroCap disposable capsule filter for concentration of coliphage MS2 and poliovirus (PV1) from deionised (DI) water and artificial seawater, as well as natural ground, surface, and seawater. Retention and recoveries for the ViroCap were compared with two well-characterised filters: the 1MDS for DI water, and the OptiCap XL for artificial seawater. The mean adsorption for MS2 by the ViroCap was 88%. Recovery of MS2 was significantly greater (p ≤ 0.01) than alternative filters tested: 65% from DI water and 63% from artificial seawater, compared to 30% for the 1MDS and 15% for the OptiCap for the respective matrices. Recovery of PV1 from DI water (37%) was similar to that of the 1MDS (51%). PV1 recoveries from artificial seawater were significantly greater (p ≤ 0.01) for the ViroCap (44%) than the OptiCap (11%). Recovery of MS2 from seeded environmental samples yielded 44% from groundwater, 53% from surface water, and 51% from seawater. ViroCap disposable filter is efficient for concentrating MS2 and PV1 from diverse matrices and is robust across a range of ionic concentrations.


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