AWWA WQTC71479

The objectives of this research included: optimization of oocyst removal from U.S. Environmental Protection Agency (USEPA) Method 1622/23 slides and DNA extraction; development of a PCR-based method for detection of human infectious and animal related Cryptosporidium genotypes; and, round-robin laboratory evaluation of the developed method. The use of bacteriological loops and closed cell foam swabs were compared for their effectiveness in scraping slide surfaces. Several different types of oocyst lysis and DNA extraction methods were evaluated. For the detection and genotyping of oocysts, a single round multiplex PCR targeting the Cryptosporidium genes for 18S ribosomal RNA (18S rDNA) and heat shock protein 70 (hsp70) was developed. Includes extended abstract only.