The Utilization of Tangential Flow Ultrafiltration and qPCR/RT-qPCR for the Simultaneous Collection and Analysis of Source Waters for Pathogenic Free Living Amoebae and Enteric Viruses

摘要

Tangential flow ultrafiltration, (TFU), was incorporated by the Water QualityrnLaboratory of the Southern Nevada Water Authority to monitor various water samplesrnfor pathogenic microorganisms. The use of TFU allows the laboratory to combine threernsampling events, virus, bacteria and amoeba, into a single sampling event. The TFUrncollected samples were then analyzed by qPCR or RT-qPCR for enteric viruses andrnpathogenic free living amoeba (PFLA). The detection target objective of 10 PFU/L forrncoxsackievirus B3, human rotavirus and adenovirus 40 was achieved. The detectionrntarget objective of 10 cells/L for Naegleria fowleri, Acanthamoeba polyphaga andrnBalamuthia mandrillaris was achieved. Sampling and analysis of environmental watersrninfluencing SNWA’s potable water source by TFU/qPCR/RT-qPCR occurred over a twornyear period. Both enteric viruses and pathogenic free living amoebae, (PFLA), wererndetected during this time period.