Hashemi FH, Di Giovanni GD, Abbaszadegan M, Lechevallier MW; American Society for Microbiology. General Meeting.
Abstr Gen Meet Am Soc Microbiol. 1999 May 30-Jun 3; 99: 549 (abstract no. Q-81).
American Water Works Service Co., Belleville, IL.
Cryptosporidium parvum is a waterborne parasite with low dose infectivity, and great health risks especially for immunocompromised individuals. Environmental grab water samples (121 raw and 122 filter backwash) from twenty five sites in the United States were collected. Approximately 10 L of each sample was concentrated by centrifugation and oocysts were recovered from the water concentrates using immunomagnetic separation (IMS). IMS was performed on up to 1.5 ml packed pellet water concentrate for each sample. Infectivity of the recovered oocysts was determined using integrated cell culture-PCR (CC-PCR). Human intestinal (HCT- 8) cells were seeded into 96-well microtiter plates. After 24 h, confluent cell monolayers were inoculated with the recovered oocysts. The plates were incubated at 37 degrees C for 72 h to allow development of infection, Infected HCT-8 cells were detected by PCR using primers specific for the heat shock protein 70 gene (hsp70) of C. parvum. CC-PCR detected infectious C. parvum in 6 raw and 9 filter backwash water samples. All CC-PCR positive samples were confirmed by cloning and DNA sequence analysis of the hsp70 PCR products. Sequence analysis revealed greater than 98% homology to the C. parvum hsp70 gene, as well as reproducible nucleotide substitutions among the infectious C parvum isolates. Results suggest that these hsp70 genotypes may represent distinct strains of C. parvum.
Publication Types:
Keywords:
- Animals
- Cryptosporidium parvum
- HSP70 Heat-Shock Proteins
- Humans
- Immunomagnetic Separation
- Oligonucleotide Probes
- Oocysts
- Polymerase Chain Reaction
- Sequence Analysis
- United States
- Water
- Water Microbiology
- Water Supply
Other ID:
UI: 102195737
From Meeting Abstracts