90-Kilodalton Heat Shock Protein, Hsp90, as a Target for Genotyping Cryptosporidium spp. Known To Infect Humans

Abstract

Small-subunit (SSU) rRNA-based methods have been commonly used in the differentiation of Cryptosporidium species or genotypes. In order to develop a new tool for confirming the genotypes of Cryptosporidium species, parts of the 90-kDa heat shock protein (Hsp90) genes of seven Cryptosporidium 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 of C. felis from cats and C. baileyi from birds, the Hsp90 genes of all tested Cryptosporidium species were amplified. Phylogenetic analysis of the hsp90 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 of hsp90 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 of Cryptosporidium in clinical specimens.

Laboratory utility of coproscopy, copro immunoassays and copro nPCR assay targeting Hsp90 gene for detection of Cryptosporidium in children, Cairo, Egypt

“…The reaction mixture consisted of 12.5 ll master mix, 200 nM from each primer, 0.2 units of Taq polymerase (as an activator) and 3 ll of the template DNA for the primary reaction and 1 ll for the secondary reaction in a total volume of 25 ll. The cycling conditions were carried out according to Feng et al (2009) with modification in the form of 50°C annealing temperature for the primary and the secondary reactions. The amplified products were visualized with 1.5 % agarose gel electrophoresis after ethidium bromide staining.…”

Genotypic Prevalence of Cryptosporidium in Egyptian Patients with Liver Cirrhosis

“…In Egypt, the predominant species in human are Cryptosporidium hominis and Cryptosporidium parvum with variance in the predominance of one more than the other in different study situations. [21][22][23] The Cryptosporidium prevalence is widely varied between countries and time. Reports showed that the Cryptosporidium prevalence is usually higher when molecular methods were used.…”

A member of the HSP90 family from ovineBabesiain China: molecular characterization, phylogenetic analysis and antigenicity

“…According to their homology, function and size, they can be divided into different families, e.g. It has also been implicated as molecular marker for identifying and differentiating parasite species and genotypes, such as Cryptosporidium and Ditylenchus (Feng et al 2009;Vovlas et al 2015). The molecular chaperone HSP90 is important in the folding and functioning of many proteins involved in cell survival, especially those participating in cell cycle regulation and signal transduction (Pearl and Prodromou, 2006).…”

“…It has also been implicated as molecular marker for identifying and differentiating parasite species and genotypes, such as Cryptosporidium and Ditylenchus (Feng et al . 2009; Vovlas et al . 2015).…”