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Applied and Environmental Microbiology, February 2003, p. 971-979, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.971-979.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Comparison of Method 1623 and Cell Culture-PCR for Detection of Cryptosporidium spp. in Source Waters

Mark W. LeChevallier,1* George D. Di Giovanni,1,{dagger} Jennifer L. Clancy,2 Zia Bukhari,2,{ddagger} Shan Bukhari,2 Jeffrey S. Rosen,3 Jose Sobrinho,3 and Michelle M. Frey4

American Water Works Service Company, Inc., Voorhees, New Jersey 08043,1 Clancy Environmental Consultants, St. Albans, Vermont 05478,2 Technology Planning & Management Consultants, Scituate, Massachusetts 02066,3 McGuire Environmental Consultants, Denver, Colorado 802104

Received 1 August 2002/ Accepted 7 November 2002

Analysis of Cryptosporidium occurrence in six watersheds by method 1623 and the integrated cell culture-PCR (CC-PCR) technique provided an opportunity to evaluate these two methods. The average recovery efficiencies were 58.5% for the CC-PCR technique and 72% for method 1623, but the values were not significantly different (P = 0.06). Cryptosporidium oocysts were detected in 60 of 593 samples (10.1%) by method 1623. Infectious oocysts were detected in 22 of 560 samples (3.9%) by the CC-PCR technique. There was 87% agreement between the total numbers of samples positive as determined by method 1623 and CC-PCR for four of the sites. The other two sites had 16.3 and 24% correspondence between the methods. Infectious oocysts were detected in all of the watersheds. Overall, approximately 37% of the Cryptosporidium oocysts detected by the immunofluorescence method were viable and infectious. DNA sequence analysis of the Cryptosporidium parvum isolates detected by CC-PCR showed the presence of both the bovine and human genotypes. More than 90% of the C. parvum isolates were identified as having the bovine or bovine-like genotype. The estimates of the concentrations of infectious Cryptosporidium and the resulting daily and annual risks of infection compared well for the two methods. The results suggest that most surface water systems would require, on average, a 3-log reduction in source water Cryptosporidium levels to meet potable water goals.

* Corresponding author. Mailing address: American Water Works Service Company, Inc., 1025 Laurel Oak Rd., P.O. Box 1770, Voorhees, NJ 08043. Phone: (856) 346-8261. Fax: (856) 782-3603. E-mail: mlecheva{at}amwater.com.

{dagger} Present address: The Texas A&M University System, El Paso, TX 79927-5020.

{ddagger} Present address: American Water Works Service Company, Inc., Belleville, IL 62221.

Applied and Environmental Microbiology, February 2003, p. 971-979, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.971-979.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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