Open Access90-Kilodalton Heat Shock Protein, Hsp90, as a Target for Genotyping Cryptosporidium spp. Known To Infect Humans
Author(s) -
Yaoyu Feng,
Theresa Dearen,
Vitaliano Cama,
Lihua Xiao
Publication year - 2009
Publication title -
eukaryotic cell
Language(s) - English
Resource type - Journals
eISSN - 1535-9778
pISSN - 1535-9786
DOI - 10.1128/ec.00294-08
Subject(s) - biology , cryptosporidium , heat shock protein , genotype , phylogenetic tree , genotyping , gene , ribosomal rna , microbiology and biotechnology , parasite hosting , genetics , feces , world wide web , computer science
Small-subunit (SSU) rRNA-based methods have been commonly used in the differentiation ofCryptosporidium species or genotypes. In order to develop a new tool for confirming the genotypes ofCryptosporidium species, parts of the 90-kDa heat shock protein (Hsp90) genes of sevenCryptosporidium species and genotypes known to infect humans (C. hominis ,C. parvum ,C. meleagridis ,C. canis ,C. muris ,C. suis , and the cervine genotype), together with one from cattle (C. andersoni ), were sequenced and analyzed. With the exception ofC. felis from cats andC. baileyi from birds, the Hsp90 genes of all testedCryptosporidium species were amplified. Phylogenetic analysis of thehsp90 sequences from all these species is congruent with previous studies in which the SSU rRNA, 70-kDa heat shock protein, oocyst wall protein, and actin genes were analyzed and showed that gastric and intestinal parasites segregate into two distinct clades. In this study, the secondary products ofhsp90 produced after PCR-restriction fragment length digestion with StyI and HphI or with BbsI showed that parasites within the intestinal or gastric clade could be differentiated from each other. These data confirm the utility of the Hsp90 gene as a sensitive, specific, and robust molecular tool for differentiating species and/or genotypes ofCryptosporidium in clinical specimens.