Article Abstract:
A polymerase chain reaction (PCR) protocol, that involves hybridization with a chemiluminescent probe, detects the oocysts of the parasite Cryptosporidium in environmental samples such as wastewater, surface water, and drinking water. The method is faster and more reliable than the generally used immunofluorescence method. Specimens are prepared for the PCR assay by pretreatment followed by a freeze-thaw procedure. Unlike the immunofluorescence assay, result of this PCR assay is not influenced by empty oocysts.
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Article Abstract:
The Cryptosporidium DNA recovery-PCR detection method effectively identifies the genomic DNA of the pathogenic protozoa Cryptosporidium in bovine feces. The test is more sensitive than the commercially used Color Vue-Cryptosporidium EIA test. The DNA is extracted by freezing and thawing dilute feces solutions followed by incubation and centrifugation. The PCR amplification involves the hybridization of the DNA with an internal oligonucleotide probe. No harmful wastes are produced during the procedure.
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Article Abstract:
Polymerase chain reaction (PCR) amplifies a gene encoding a repetitive oocyst protein from a DNA fragment of Cryptosporidium parvum. The oligonucleotide primers used in the PCR analysis are based on the amino acid sequence of the oocyte protein gene. Live and dead oocytes are discriminated by an excystation protocol before the DNA extraction. Nested PCR enhance sensitivity which allows the detection of viable C. parvum present in small amounts.
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