Journal article

Detection of a single viable Cryptosporidium parvum oocyst in environmental water concentrates by reverse transcription-PCR

T Stinear, A Matusan, K Hines, M Sandery

Applied and Environmental Microbiology | AMER SOC MICROBIOLOGY | Published : 1996

Abstract

Current methods for detection of Cryptosporidium parvum oocysts in water are time-consuming and difficult. We have developed a reverse transcription (RT)-PCR which can detect the presence of a single viable oocyst spiked into concentrated environmental water samples. The test is based on the detection of mRNA from a C. parvum heat shock protein (hsp). The synthesis of hsp was induced by a short 45 degrees C incubation followed by oocyst lysis by a freeze-thaw process. Hsp70 mRNA, produced only from viable oocysts, was then isolated by hybridization to oligo(dT)25-coated magnetic beads. Detection was achieved by RT-PCR amplification of a 590-bp region of hsp70 mRNA specific for C. parvum. To ..

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