Full Description

The main goal of this study was to develop a molecular method alternative to the currenttechnique, based on immunofluorescence, with the same detection limits, high specificity andbe more secure. This protocol must permit making confident and comfortable decisions incases of Cryptosporidium incidents.This study presents the first results on the development, including the design of probe andprimers with HSP 70 gene as target, the comparison and optimization of two DNA extractionmethods. To build the standard curve, the study prepared a suspension of oocysts purified fromfaeces, made a numeration with a solid phase cytometer and performed a DNA extraction toprepare a stock solution.With this protocol, the study could detect up to 2 oocysts. The detection of 1 copy is achieved in60% of the cases.Currently, the study needs to validate this approach on real samples. Preliminary results arepromising.Availability of such a tool would allow multiple controls at the different stages of thewater production and possibly the analysis of more complex matrices or coupling thisprotocol with an infectivity protocol. Includes 6 references, tables, figure.

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Edition: Vol. - No. Published: 11/02/2003 Number of Pages: 8File Size: 1 file , 270 KB