Quantitative RT-PCR Detection of Hepatitis A Virus, Rotaviruses and Enteroviruses in the Buffalo River and Source Water Dams in the Eastern Cape Province of South Africa

Abstract

Human enteric viruses (HEntVs) are a major cause of water-related diseases. The prevalence of hepatitis A virus (HAV), rotaviruses (RoV) and enteroviruses (EnV) in Buffalo River waters was assessed quantitatively over a period of 12 months (August 2010 to July 2011). Seventy-two samples were collected from six sites, including three dams, and concentrated using the adsorption-elution method. Viral RNA was extracted using a commercial kit, and the viruses were quantified by real-time quantitative reverse transcriptase PCR (RT-qPCR). Two or more viruses were detected in 12.5% of the samples. HAV was detected in 43.1% of the samples and in significantly (p < 0.05) varying concentrations of 1.5 × 101–1.9 × 105 genome copies/L compared to RoV and EnV, while RoVs were detected in 13.9% of samples, with concentrations ranging from 2.5 × 101–2.1 × 103 genome copies/L, and EnV were detected in 9.7% of the samples, with concentrations ranging from 1.3 × 101–8.6 × 101 genome copies/L. Only HAV was detected at all the sites, with the Bridle Drift Dam recording significantly higher (p < 0.05) concentrations. The presence of enteric viruses in Buffalo River may constitute public health risks and the incidence of HAV at all the sites could reflect both the epidemiological status of hepatitis A and HAV persistence in the water environments.

Monitoring of Foodborne Hepatitis A virus Outbreaks in the Fresh Foods

“…Additionally, HAV tends to be labile and loss of infectivity can occur during transport. Many studies have reported that nucleic acid amplification technique are more sensitive than viral culture for detecting HAV in samples (Chigor & Okoh, 2012). This may be explained at least in part by non-viability of viral particles in the specimens that can be detect by RT-PCR was used for HAV detection.…”

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

“…Centrifugal ultrafiltration removes water and components less than 30,000 Daltons resulting in both concentration of any virus in test samples and a reduction in small molecular weight inhibitors of molecular assays. This tertiary concentration step results in an overall concentration factor of >10 5 for any virus that was present in the water being tested.…”

“…Both tools have demonstrated that many virus types are present in environmental and drinking waters throughout the world [1][2][3][4][5][6] . Their use coupled with sequencing of amplified genomic fragments during disease outbreak investigations has provided evidence for waterborne virus transmission, as they have shown that the virus found in the drinking water is identical to that shed by outbreak patients [7][8][9][10] .…”

Infección por el virus de la hepatitis A: epidemiología y diversidad genética

“…Las partículas del VHA son estables en el medio ambiente, permanecen de días a meses en suelos, fuentes de agua, sedimento marino y ostras (5,(26)(27)(28)(29)(30). Son resistentes al pH en un rango de 3 a 10; se requiere un pH superior a 10 para inactivarlas.…”

“…Las regiones del Sureste de Asia y África Subsahariana presentan una alta endemicidad (14,26,40); el Mediterráneo Oriental, Asia Central, África del Norte, Centroamérica y Suramérica presentan endemicidad intermedia (1,14). Europa Central y Oriental presentan baja endemicidad mientras que Europa Occidental, Norteamérica y la región del Suroeste de Oceanía presentan muy baja endemicidad (38).…”