Hepatitis E virus in lettuce and water samples: A method-comparison study

Randazzo, Walter; Vásquez-García, Andrea; Bracho, María Alma; Alcaráz, María Jesús; Aznar, Rosa; Sánchez, Glòria

doi:10.1016/j.ijfoodmicro.2018.04.008

Cited by 14 publications

(12 citation statements)

References 39 publications

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“…Initially, two different RT-qPCRs were applied to assess HEV occurrence. Surprisingly, RT-qPCR1 showed a limited number of positives compared to RT-qPCR2 ( Figure 5) despite a previous study reported similar performance of these assays in influent water samples (Randazzo et al, 2018). Suspecting that a different HEV genotype was circulating, a third RT-qPCR assay was included in the study.…”

Section: Performance Of Rt-qpcr Assays Of Hev In Naturally Contaminatmentioning

confidence: 95%

HEV Occurrence in Waste and Drinking Water Treatment Plants

et al. 2020

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Hepatitis E virus (HEV), particularly zoonotic genotype 3, is present in environmental waters worldwide, especially in industrialized countries. Thus, monitoring the presence of HEV in wastewater treatment plants (WWTPs) is an emerging topic due to the importance of reusing water on a global level. Given the limited data, this study aimed to monitor the occurrence of HEV in influent and effluent water in waste-and drinkingwater treatment plants (WWTPs and DWTPs). To this end, different procedures to concentrate HEV in influent and effluent water from WWTPs and DWTPs were initially evaluated. The evaluated procedures resulted in average HEV recoveries of 15.2, 19.9, and 16.9% in influent, effluent, and drinking water samples, respectively, with detection limits ranging from 10 3 to 10 4 international units (IU)/L. Then, a one-year pilot study was performed to evaluate the performance of the selected concentration method coupled with three RT-qPCR assays in influent and effluent water samples from four different WWTPs. HEV prevalence in influent water varied based on both the RT-qPCR assay and WWTP, while HEV was not detected in effluent water samples. In addition, HEV prevalence using only RT-qPCR3 was evaluated in influent (n = 62) and effluent samples (n = 52) from four WWTPs as well as influent (n = 28) and effluent (n = 28) waters from two DWTPs. The present study demonstrated that HEV circulated in the Valencian region at around 30.65% with average concentrations of 6.3 × 10 3 IU/L. HEV was only detected in influent wastewater samples, effluent samples from WWTPs and influent and effluent samples from DWTPs were negative. However, given that the infective dose in waterborne epidemics settings is not yet known and the low sensibility of the assay, unfortunately, no direct conclusion could be achieved on the risk assessment of environmental contamination.

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Section: Performance Of Rt-qpcr Assays Of Hev In Naturally Contaminatmentioning

confidence: 95%

HEV Occurrence in Waste and Drinking Water Treatment Plants

et al. 2020

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“…HEV is transmitted mainly by the consumption of contaminated water or contaminated undercooked or raw food such as vegetables, meat (pork, mutton, rabbit, poultry) and dairy products [2,5,14,15]. It is important to note that contaminated water is not only a cause of direct transmission but is also related to indirect transmission due to its use in agriculture practices, namely in the irrigation of vegetables, where the virus may accumulate and be delivered as infectious particles [16][17][18][19][20]. Although less frequent, HEV can also be transmitted by transfusions with contaminated blood and by vertical (mother-to-child) transmission [14,21,22].…”

Section: Introductionmentioning

confidence: 99%

Assessment of the Presence of Hepatitis E Virus in Surface Water and Drinking Water in Portugal

et al. 2020

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Hepatitis E virus (HEV) is a non-enveloped single-stranded positive-sense RNA virus, belonging to the Hepeviridae family, resistant to environmental conditions, and transmitted by the consumption of contaminated water. This virus is responsible for both sporadic and epidemic outbreaks, leading to thousands of infections per year in several countries, and is thus considered an emerging disease in Europe and Asia. This study refers to a survey in Portugal during 2019, targeting the detection and eventual quantification of enteric viruses in samples from surface and drinking water. Samples positive for HEV RNA were recurrently found by reverse transcription quantitative PCR (RT-qPCR), in both types of matrix. The infectivity of these samples was evaluated in cultured Vero E6 cells and RNA from putative viruses produced in cultures evidencing cytopathic effects and was subjected to RT-qPCR targeting HEV genomic RNA. Our results evidenced the existence of samples positive either for HEV RNA (77.8% in surface water and 66.7% in drinking water) or for infectious HEV (23.0% in surface water and 27.7% in drinking water). These results highlight the need for effective virological control of water for human consumption and activities.

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“…In this study, three previously described HEV assays (Mansuy et al, 2004; Randazzo et al, 2018c) were coupled with two viability markers propidium monoazide (PMAxx) and platinum chloride (PtCl 4 ) and initially evaluated on purified viral RNA. The optimized viability RT-qPCR method was then applied to native, heat-, and high-pressure processing (HPP)-treated HEV to assess its performance in discriminating between potentially infectious and inactivated viral particles.…”

Section: Introductionmentioning

confidence: 99%

Viability RT-qPCR to Distinguish Between HEV and HAV With Intact and Altered Capsids

et al. 2018

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The hepatitis E virus (HEV) is an emerging pathogen showing a considerable increase in the number of reported cases in Europe mainly related to the ingestion of contaminated food. As with other relevant viral foodborne pathogens, real-time reverse transcriptase polymerase chain reaction (RT-qPCR) is the gold standard for HEV detection in clinical, food, and environmental samples, but these procedures cannot discriminate between inactivated and potentially infectious viruses. Thus, the aim of this study was to develop a viability PCR method to discriminate between native, heat-, and high-pressure processing (HPP)-treated HEV using the hepatitis A virus (HAV) as a cultivable surrogate. To this end, different concentrations of viability markers (PMAxx and platinum chloride, PtCl4) were screened firstly on purified viral RNA using different RT-qPCR assays. Reductions of HEV RNA signals of >17.5, >15.0, and >15.5 quantification cycles (Cq) were reported for PtCl4 and 1.6, 2.9, and 8.4 Cq for PMAxx, clearly indicating a better performance of PtCl4 than PMAxx irrespective of the RT-qPCR assay used. The most efficient viability pretreatment (500 μM PtCl4 incubated at 5°C for 30 min) was then assessed on native, heat-, and HPP-treated HEV suspension. The optimized viability RT-qPCR discriminated successfully between native, heat-, and HPP-treated HEV, to different extents depending on the experimental conditions. In particular, approximately 2-log10 reduction was reported by PtCl4-RT-qPCR at both 72 and 95°C compared to the control. Additionally, both viability pretreatments were tested for HPP-treated HAV without success, while PtCl4-RT-qPCR completely eliminated (>5.6-log10 reduction) the RT-qPCR signals of HPP-treated HEV. Although this viability procedure may still overestimate infectivity, the PtCl4 pretreatment represents progress to better interpreting the quantification of intact HEV, and it could be included in molecular procedures used to quantify enteric viruses in food and environmental samples.

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Hepatitis E virus in lettuce and water samples: A method-comparison study

Cited by 14 publications

References 39 publications

HEV Occurrence in Waste and Drinking Water Treatment Plants

HEV Occurrence in Waste and Drinking Water Treatment Plants

Assessment of the Presence of Hepatitis E Virus in Surface Water and Drinking Water in Portugal

Viability RT-qPCR to Distinguish Between HEV and HAV With Intact and Altered Capsids

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