RESUMEN

We describe a membrane-filter-based urea-arginine phosphate buffer method for concentrating waterborne viruses from large volumes of water to microlitre volumes, and their subsequent detection by the polymerase chain reaction (PCR). The detection step involves the extraction of RNA, synthesis of complementary DNA, amplification by PCR of target DNA with specific primers, and confirmation through nucleic acid hybridization with a radiolabelled oligonucleotide probe. The PCR technique detected the presence of enteroviruses in spiked as well as in contaminated water samples. The technique is sensitive and detects as few as 120 waterborne viral particles. PCR is simple, rapid, sensitive, specific and adaptable for water quality surveillance in less developed countries.

Asunto(s)

Poliovirus/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Microbiología del Agua , Secuencia de Bases , Southern Blotting , Colodión , Filtración/métodos , Genes Virales/genética , Membranas Artificiales , Datos de Secuencia Molecular , Poliovirus/genética , ARN Viral/aislamiento & purificación , Sensibilidad y Especificidad , Proteínas no Estructurales Virales/genética