摘要
Three isolates of the genus Cryptosporidium, namely, Guangdong isolate, Anhui isolate and Jiangsu isolate from MainlandChina, were identified and characterized genetically utilizing nuclear DNA regions of the small subunit of ribosomal RNA(SSU rRNA) and heat shock protein 70 gene (HSP70) as genetic markers. These two regions were amplified by PCR fromDNA extracted from oocysts and amplicons of approximately 290 bp and 450 bp were produced, respectively. The ampliconswere purified, cloned and sequenced. Sequences of 446 bp and 290-292 bp were obtained for the SSU rRNA and HSP70regions, respectively. The obtained SSU rRNA and HSP70 sequences representing the three Cryptosporidium isolateswere compared with those retrieved from the DNA database. Genetic analyses using either DNA region revealed thatmembers of Cryptosporidium formed two clusters, with C. parvum, C. wariri, C. felis and C. meleagridis clusteredtogether, while C. andersoni, C. muris and C. serpentis belong to the other cluster. Based on SSU rRNA and HSP70sequences, both Guangdong and Anhui isolates of Cryptosporidium were identified as C. muris of the calf genotype (i.e., C. andersoni), whereas the Jiangsu isolate was identified as C. parvum of the calf genotype. The findings of thepresent study should have important implications for the diagnosis and control of Cryptosporidium infections in bothhumans and animals in China.
Three isolates of the genus Cryptosporidium, namely, Guangdong isolate, Anhui isolate and Jiangsu isolate from MainlandChina, were identified and characterized genetically utilizing nuclear DNA regions of the small subunit of ribosomal RNA(SSU rRNA) and heat shock protein 70 gene (HSP70) as genetic markers. These two regions were amplified by PCR fromDNA extracted from oocysts and amplicons of approximately 290 bp and 450 bp were produced, respectively. The ampliconswere purified, cloned and sequenced. Sequences of 446 bp and 290-292 bp were obtained for the SSU rRNA and HSP70regions, respectively. The obtained SSU rRNA and HSP70 sequences representing the three Cryptosporidium isolateswere compared with those retrieved from the DNA database. Genetic analyses using either DNA region revealed thatmembers of Cryptosporidium formed two clusters, with C. parvum, C. wariri, C. felis and C. meleagridis clusteredtogether, while C. andersoni, C. muris and C. serpentis belong to the other cluster. Based on SSU rRNA and HSP70sequences, both Guangdong and Anhui isolates of Cryptosporidium were identified as C. muris of the calf genotype (i.e., C. andersoni), whereas the Jiangsu isolate was identified as C. parvum of the calf genotype. The findings of thepresent study should have important implications for the diagnosis and control of Cryptosporidium infections in bothhumans and animals in China.
基金
Financial support was provided by the Natural Science Foundation of Guangdong Province(010354,32286)
China and National Natural Science Foundation of China(30371082).
