Molecular Detection and Genotyping of Group A Rotavirus by Multiplex Semi-Nested RT-PCR in Sewage Water and Sludge

Group A rotavirus (RV-A) infections are a major cause of morbidity and mortality in human around the world. The aim of this research was to conduct a molecular characterization of RV-A glycoprotein (G) and protease-sensitive (P) in urban sewage (n=54; 27 raw sewage and 27 treated sewage) and sewage sludge (n=27) from Egypt by multiplex semi-nested RT-PCR method. RV-A was detected in 29.9% of raw sewage, 7.4% of treated sewage, and 18.5% of sewage sludge samples. In the positive samples, the RV-A G genotypes were as follow: G3 (n=6), G9 (n=3), G1 (n=2), G4 (n=1), G10 (n=1), G1+G3 (n =1), and G1+G3+G10 (n=1) whereas P types detected included: P[4] (n=1), P[6] (n=3), P[8] (n=8), and nontypeable P (n=3). The most detected G-P combination was G3P [8] (n=6). Other G-P combinations such as G1P [8] (n=2), G9P [6] (n=2), G4P [6] (n=1), and G10P[4] (n=1) were also detected. The highest detection rates of RV-A in sewage samples were found in winter (50%), followed by summer (25%), then in spring and autumn (12.5%). The results showed not only that sewage analysis contains dramatic information concerning enteric viruses, but also that environmental monitoring is an important approach in describing the local circulation of specific viruses among population.