Quantitative RT-PCR detection of human noroviruses and hepatitis A virus in fresh produce and surface water used for irrigation in the Mansoura and Giza regions, Egypt

Author(s):  
Mohamed N. F. Shaheen ◽  
Elmahdy M. Elmahdy ◽  
Lamiaa H. I. Mahmoud ◽  
Ibtisam A. Hammad ◽  
Elham R. S. Soliman
2021 ◽  
Author(s):  
Mohamed N.F. Shaheen ◽  
Elmahdy M. Elmahdy ◽  
Lamiaa H. I. Mahmoud ◽  
Ibtisam A. Hammad ◽  
Elham R.S. Soliman

Abstract Vegetables can be considered as a vehicle for the transmission of human enteric viruses such as noroviruses (genogroups I and II) and hepatitis A virus (HAV) when irrigated with contaminated irrigation or when prepared by infected food handlers. In the current study, we investigated the presence of hepatitis A virus (HAV) and human noroviruses (genogroups I and II) in fresh produce and surface water used in cultivation of this produce using real-time PCR. Samples were collected from six different points in the Mansoura and Giza regions, Egypt. Our analysis showed that at least one virus was found in 41.6% (30/72) of surface water samples and 27% (13/48) of fresh produce samples. HAV (23/72) with a mean viral concentration = 4 × 106 genome copies/litter (GC/L) was the most frequently identified virus in surface irrigation water samples, followed by human norovirus genogroup II (HNoV GII) (15/72, with a mean concentration = 1.2 × 106 GC/L, and human noroviruses genogroup I (HNoV GI) (12/72, with a mean concentration = 1.4 × 104 GC/L). Additionally, HAV (10/48) with a mean concentration = 5.2 × 105 genome copies/gram (GC/g) was also the most frequently detected virus in the fresh produce samples, followed by HNoV GII (8/48, with a mean concentration = 1.7 × 104 GC/g), meanwhile HNoV GI (6/48) was less detected virus with a mean concentration = 3 × 103 GC/g. This work suggests a wide prevalence of human enteric viruses in surface irrigation waters and fresh produce, which is of concern when the fresh produce is eaten raw. Thus, Additional monitoring for viral pathogens in irrigation water and food is needed to increase produce safely.


2004 ◽  
Vol 50 (1) ◽  
pp. 131-136 ◽  
Author(s):  
J.L. Romalde ◽  
C. Ribao ◽  
M. Luz Vilariño ◽  
J.L. Barja

In the present study, the efficiency of several primer sets for the RT-PCR detection of hepatitis A virus (HAV) and astrovirus from both crude viral extracts and experimentally infected shellfish tissues was evaluated. Differences were observed depending on the primer set employed in the sensitivity of amplification of both viral types. For HAV primers, HAV240/HAV68 yielded the higher sensitivity: showing a detection limit of 0.02-0.1 infectious particles/μL or mg of tissue (either crude extracts or seeded mussel tissues). Regarding detection of AsV, a better performance was observed with primer set A1/A2 achieving a sensitivity of 0.1-1 PFU/μL or mg of tissue. The results obtained in this work strongly indicated that selection of primer sets to be employed for the routine detection of enteric viruses was a critical point in the design of the RT-PCR protocols.


2009 ◽  
Vol 157 (1) ◽  
pp. 80-83 ◽  
Author(s):  
Sylvie Perelle ◽  
Laetitia Cavellini ◽  
Christian Burger ◽  
Sandra Blaise-Boisseau ◽  
Catherine Hennechart-Collette ◽  
...  

1991 ◽  
Vol 24 (2) ◽  
pp. 267-272 ◽  
Author(s):  
S. Dubrou ◽  
H. Kopecka ◽  
J. M. Lopez Pila ◽  
J. Maréchal ◽  
J. Prévot

Enteroviruses were specifically detected by dot blot hybridization when using poliovirus type 1 (PV1) derived subgenomic radiolabeled cRNA probes (riboprobes) in environmental water specimens and in the cell cultures in which the viruses were amplificated. The riboprobe corresponding to the 5' noncoding sequence detected the majority of enteroviruses. Hepatitis A virus (HAV) was specifically detected by an HAV cRNA probe corresponding to the 5' noncoding region of its genome. By this test, the limit of detection of coxsackievirus B5 and echovirus 7 seeded in mineral water was 103 to 104 PFU/spot. In cell cultures, positive signals were observed in the lysates of cells infected by one PFU. Higher positive signals were obtained with a short PV1 probe (nt 221-670) corresponding to the 5' noncoding region, which is a well preserved sequence among the enteroviruses, than with PV1 genomic probe. Hybridization allowed a good detection of enteroviral RNAs in wastewater specimens, but with a lower efficiency in surface water. In this case, amplification of viruses in the cell cultures gave significant hybridization results.


2017 ◽  
Vol 9 (4) ◽  
pp. 395-405 ◽  
Author(s):  
Emmy Borgmästars ◽  
Mehrdad Mousavi Jazi ◽  
Sofia Persson ◽  
Linda Jansson ◽  
Peter Rådström ◽  
...  

2015 ◽  
Vol 41 (04) ◽  
pp. 229-235
Author(s):  
Kuang-Po Li ◽  
Shan-Chia Ou ◽  
Jui-Hung Shien ◽  
Poa-Chun Chang

Duck hepatitis A virus type 1 (DHAV-1) infection is a highly contagious and fatal disease of young ducklings. A live attenuated vaccine strain designated as 5886 has been used in Taiwan for the control of DHAV-1. Although several molecular biological methods are reported for diagnosis of DHAV-1 infection, none of them is able to discriminate between the vaccine strain and field viruses of DHAV-1. In the present study, a real-time reverse transcriptase polymerase chain reaction (RT-PCR) and high resolution melting (HRM) assay was developed for rapid detection and differentiation between the vaccine strain and field viruses of DHAV-1. This assay is highly specific for DHAV-1 and the detection limit is about 100 copies of the viral RNA. Experiments using fecal samples collected from ducklings experimentally infected with DHAV-1 showed that DHAV-1 could be detected in fecal samples as early as 6 h post-infection. In summary, a real-time RT-PCR and HRM assay is developed in this study and this assay could be valuable for diagnosis and surveillance of DHAV-1 infection in the field.


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