Sub-species typing among bovine C. parvum isolates by PCR-PAGE using a novel microsatellite + telomere primer scheme

Abstract

A PCR-PAGE procedure based on a novel primer set including a previously described microsatellite (M) plus a telomere anchor (+T) was used to biotype C. parvum isolated from bovine sources in three categories: geographically diverse, intra herd, and specifically defined. The PAGE patterns resulting from the procedure consisted of from 8 to 18 bands in the 300 to 3,000 bp size range. Application of the procedure revealed widely divergent PAGE banding among the geographically diverse samples. Consistent banding patterns were observed among multiple calf hosts from the same dairy herd sampled over 1 to 2 year periods. Some differences were observed in PAGE banding between samples in different time periods although all independent samples in each sampling period (up to 5 shown) were homogeneous. Analysis of DNA from samples of the Iowa defined strain of C. parvum isolated in 1998 and in 2000 from the same source (University of Arizona) resulted in PAGE banding that was homogeneous. The sub-type differentiation effectiveness of the M+T PAGE procedure was observed to be substantial. Reference to work being prepared for publication elsewhere indicates that it is similarly effective for Type 1 samples.