Article Abstract:
An technique has been devised to detect low-levels of infective Cryptosporidium oocysts using HCT-8 cells in the Cryptosporidium parvum reproductive cycle. An indirect-antibody assay called the focus detection method is described. It uses primary and secondary antibodies and epifluorescence microscopy to selectively identify infection and produces more reliable quantifiable results than the oocyst detection method currently employed in the water industry.
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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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Article Abstract:
High levels of iron in soil or water could interfere with methods for detecting Cryptosporidium parvum and Giardia duodenalis. The most commonly used method is immunomagnetic separation-immunofluorescent assay method, but dissolved iron reduced the recovery rate of the two organisms. This is important because iron is the second most common metal in soil.
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