Detection of Infectious Noroviruses from Wastewater and Seawater Using PEMAXTM Treatment Combined with RT-qPCR

Gyawali, Pradip; Hewitt, Joanne

doi:10.3390/w10070841

Cited by 18 publications

(15 citation statements)

References 33 publications

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“…Even if PMAxx showed to significantly affect the virus titers (Table S2), the high loads of viruses in the PMAxx-treated samples indicate the potential infectivity of detected viruses. Similarly, Gyawali and Hewitt (2018) evaluated the performance of PEMAX-RT-qPCR on influent and effluent water and found a high proportion of potentially infectious NoV GI and GII (i.e., 21 to 89% in influent, and 24 to 59% in effluent).…”

Section: Interlaboratory Study For the Detection And Quantification Of Potentially Infectious Enteric Viruses In Influent And Effluent Wamentioning

confidence: 99%

Interlaboratory Comparative Study to Detect Potentially Infectious Human Enteric Viruses in Influent and Effluent Waters

Randazzo

Piqueras²,

Evtoski

et al. 2019

Food Environ Virol

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Wastewater represents the main reusable water source after being adequately sanitized by wastewater treatment plants (WWTP). In this sense, only bacterial quality indicators are usually checked to this end, and human pathogenic viruses usually escape from both sanitization procedures and controls, posing a health risk on the use of effluent waters.In this study, we evaluated a protocol based on aluminium adsorption-precipitation to concentrate several human enteric viruses, including norovirus genogroup I (NoV GI), NoV GII, hepatitis A virus (HAV), astrovirus (HAstV), and rotavirus (RV), with limits of detection of 4.08, 4.64, 5.46 log genomic copies/L, 3.31, and 5.41 log PCR units (PCRU)/L, respectively. Furthermore, the method was applied in two independent laboratories to monitor the presence of NoV GI, NoV GII, and HAV in effluent and influent waters collected from five WWTPs at two different sampling dates.Concomitantly, a viability PMAxx-RT-qPCR was applied to all the samples to get information on the potential infectivity of both influent and effluent waters. The range of the titers in influent waters for NoV GI, NoV GII, RV and HAstV was 4.80-7.56, 5.19-7.31 log genomic copies/L, 5.41-6.52, and 4.59-7.33 log PCRU/L, respectively. In effluent waters, the titers ranged between 4. 08-6.27, 4.64-6.08 log genomic copies/L, <5.51, and 3.31-5.58 log PCRU/L. Moreover, the viral titers detected by viability RT-qPCR showed statistical differences with RT-qPCR alone, suggesting the potential viral infectivity of the samples despite some observed reductions. The proposed method could be applied in ill-equipped laboratories, due to the lack of a requirement for a specific apparatus (i.e., ultracentrifuge).

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Section: Interlaboratory Study For the Detection And Quantification Of Potentially Infectious Enteric Viruses In Influent And Effluent Wamentioning

confidence: 99%

Interlaboratory Comparative Study to Detect Potentially Infectious Human Enteric Viruses in Influent and Effluent Waters

Randazzo

Piqueras²,

Evtoski

et al. 2019

Food Environ Virol

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“…Higher temperatures, which are known to denaturate the virus capsid, showed comparable results when analyzed with azo-dyes. Articles published by [34,[67][68][69] indicate that inactivation of Noroviruses GI/GII and the Hepatitis A virus, as well as Rotavirus and Aichivirus 1 after inactivation at temperatures between 50 • C and 95 • C was observable when PMA derived PMAxx, PMA, and EMA pretreatment was applied.…”

Section: Resultsmentioning

confidence: 99%

Capsid Integrity qPCR—An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

Leifels

Shoults

Wiedemeyer

et al. 2019

Water

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Recreational, reclaimed and drinking source waters worldwide are under increasing anthropogenic pressure, and often contain waterborne enteric bacterial, protozoan, and viral pathogens originating from non-point source fecal contamination. Recently, the capsid integrity (ci)-qPCR, utilizing the azo-dyes propidium monoazide (PMA) or ethidium monoazide (EMA), has been shown to reduce false-positive signals under laboratory conditions as well as in food safety applications, thus improving the qPCR estimation of virions of public health significance. The compatibility of two widely used human adenovirus (HAdV) qPCR protocols was evaluated with the addition of a PMA/EMA pretreatment using a range of spiked and environmental samples. Stock suspensions of HAdV were inactivated using heat, UV, and chlorine before being quantified by cell culture, qPCR, and ci-qPCR. Apparent inactivation of virions was detected for heat and chlorine treated HAdV while there was no significant difference between ci-qPCR and qPCR protocols after disinfection by UV. In a follow-up comparative analysis under more complex matrix conditions, 51 surface and 24 wastewater samples pre/post UV treatment were assessed for enteric waterborne HAdV to evaluate the ability of ci-qPCR to reduce the number of false-positive results when compared to conventional qPCR and cell culture. Azo-dye pretreatment of non-UV inactivated samples was shown to improve the ability of molecular HAdV quantification by reducing signals from virions with an accessible genome, thereby increasing the relevance of qPCR results for public health purposes, particularly suited to resource-limited low and middle-income settings.

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“…The technique, which has enabled the copy and quantification of fragments of the virus' genetic material through in-vitro replications, has been considered as reliable, sensitive, highly specific and the "gold standard" in the detection of low amounts of genetic material in different types of matrices (Corpuz et al 2020;Hamouda et al 2021). Earlier studies indicated that RT-qPCR methods were successfully able to determine the diversity and abundance of several viral pathogens in wastewater, including Adenovirus, Astrovirus, Enterovirus, Hepatitis A and E, Norovirus, Papillomavirus, Parechovirus, Polyomavirus, Rotavirus and Salivirus (O'Brien et al 2017;Bisseux et al 2018;Gyawali & Hewitt 2018;Adineh et al 2019;I. Ahmed et al 2019;Randazzo et al 2019).…”

Section: Methods Used To Detect the Sars-cov-2 In Wastewatermentioning

confidence: 99%

Early detections of SARS-CoV-2 in wastewater and their use in COVID-19 epidemiological control

Mainardi

Bidóia

2021

RSD

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SARS-CoV-2 is a new type of coronavirus that is capable to infect humans and cause the severe acute respiratory syndrome COVID-19, a disease that has been causing enormous impacts across the whole Earth. COVID-19 patients, including mild, pre-symptomatic and asymptomatic cases, were often seen to contain infectious fragments of SARS-CoV-2 in urine and feces samples. Thus, studies to detect the new coronavirus in wastewater, which collect and concentrate human excreta, have been extremely useful as a tool to monitor the virus spread in the communities. The surveillance, frequently used to elaborate economical non-invasive diagnoses about the circulation of chemicals and pollutants in populations, could be used as a quick alert about emerging and reemerging COVID-19 epidemics, improve the predictions concerning the SARS-CoV-2 dissemination and promote the development of better viral containment measures. Along that, the approach could be used to construct more accurately epidemiological models, foment better practices to coordinate resources an administer the vaccines, assess the propagation of SARS-CoV-2 variants across temporal and geographic scales, evaluate environmental risks and also the effectiveness of disinfection systems. In this review, it was addressed early reports regarding SARS-CoV-2 detections in wastewater and the importance of this valuable approach in the application of faster and more effective interventions from public health authorities, either towards the COVID-19 or any other epidemic disease in the future.

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Detection of Infectious Noroviruses from Wastewater and Seawater Using PEMAXTM Treatment Combined with RT-qPCR

Cited by 18 publications

References 33 publications

Interlaboratory Comparative Study to Detect Potentially Infectious Human Enteric Viruses in Influent and Effluent Waters

Interlaboratory Comparative Study to Detect Potentially Infectious Human Enteric Viruses in Influent and Effluent Waters

Capsid Integrity qPCR—An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

Early detections of SARS-CoV-2 in wastewater and their use in COVID-19 epidemiological control

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