Full Description

The use of molecular methods, such as polymerase chain reaction (PCR), in situ hybridization or DNA chips, is very useful in order to improve the microbiological control of drinking water or wastewater. The purpose of this work was to demonstrate the feasibility of the use of a quantitative RT-PCR method (Taqman technology) in a routine laboratory using two examples: the detection of enteroviruses and hepatitis A viruses (HAV) from sludge samples. Two protocols for the detection of enteroviruses and HAV using real time quantitative RT-PCR were developed and adapted to the analysis of sludge samples in a laboratory employing very skilled specialists of molecular biology (laboratory of virology of CHU of Nantes, France). In order to use these protocols routinely, evaluation tests were performed by laboratory personal that relied on the validation of the protocol of real time quantitative RT-PCR: detection limit, repeatability, and also on the validation of the complete protocol including the concentration and purification of sludge samples and the RT-PCR detection of viruses in an interlaboratory trial. Results show the feasibility of the transfer and the use of new technologies, such as quantitative PCR, in a routine laboratory. Includes 6 references, tables.

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Edition: Vol. - No. Published: 11/01/2002 Number of Pages: 8File Size: 1 file , 260 KB