AWWA QTC98356

The objective of this research was to develop a molecular fingerprinting assay that could distinguish C. parvum from other species of Cryptosporidium and differentiate the host of origin from a variety of environmental samples. In developing a method that was sensitive and specific for a range of environmental samples (e.g., feces, source water, finished water), the freeze/thaw method was compared to the use of a commercial DNA extraction kit for soil (FastDNA[TM]). Purified oocysts were obtained from outside laboratories. Fresh fecal samples were collected and stored in 0.1-percent phosphate-buffered saline (PBS) solution. All samples were pretreated in a chlorine solution and neutralized with sodium thiosulfate. The samples were then either processed by the freeze/thaw method, as previously reported or extracted with the FastDNA kit according to the manufacturer's recommendations. C. parvum DNA was also obtained from a university laboratory. Two previously published arbitrary primers were used to randomly amplify the same template DNA from an earlier study under the same annealing conditons. The resulting banding patterns were compared to the originally published patterns. C. parvum DNA from bovine (Immunocell) and human (Waterborne) hosts of origin were also randomly amplified under the same primer and annealing conditions.

Product Details

Published:

01/01/1998

ISBN(s):

0898679796

Number of Pages:

11

File Size:

1 file , 85 KB