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• W4244931116 endingPage "1749" @default.
• W4244931116 startingPage "1743" @default.
• W4244931116 abstract "ABSTRACT We recently described a reverse transcription-PCR (RT-PCR) for detecting low numbers of viable Cryptosporidium parvum oocysts spiked into clarified environmental water concentrates. We have now modified the assay for direct analysis of primary sample concentrates with simultaneous detection of viable C. parvum oocysts, Giardia cysts, and a novel type of internal positive control (IPC). The IPC was designed to assess both efficiency of mRNA isolation and potential RT-PCR inhibition. Sensitivity testing showed that low numbers of organisms, in the range of a single viable cyst and oocyst, could be detected when spiked into 100-μl packed pellet volumes of concentrates from creek and river water samples. The RT-PCR was compared with an immunofluorescence (IF) assay by analyzing 29 nonspiked environmental water samples. Sample volumes of 20 to 1,500 liters were concentrated with a wound fiberglass cartridge filter. Frequency of detection for viable Giardia cysts increased from 24% by IF microscopy to 69% by RT-PCR. Viable C. parvum oocysts were detected only once by RT-PCR (3%) in contrast to detection of viable Cryptosporidium spp. in four samples by IF microscopy (14%), suggesting that Cryptosporidium species other than C. parvum were present in the water. This combination of the large-volume sampling method with RT-PCR represents a significant advance in terms of protozoan pathogen monitoring and in the wider application of PCR technology to this field of microbiology." @default.
• W4244931116 created "2022-05-12" @default.
• W4244931116 creator A5028204622 @default.
• W4244931116 creator A5035786218 @default.
• W4244931116 date "1998-05-01" @default.
• W4244931116 modified "2023-10-14" @default.
• W4244931116 title "Sensitive and Rapid Detection of Viable <i>Giardia</i> Cysts and <i>Cryptosporidium parvum</i> Oocysts in Large-Volume Water Samples with Wound Fiberglass Cartridge Filters and Reverse Transcription-PCR" @default.
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• W4244931116 doi "https://doi.org/10.1128/aem.64.5.1743-1749.1998" @default.
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