摘要
从河南两个地区猪的粪便中分离纯化了猪源隐孢子虫卵囊。参考隐孢子虫Hsp70基因属特异性引物，用PCR分别扩增了卵囊基因组DNA大小均为1948bp的片段，PCR产物经电泳鉴定后用试剂盒回收纯化，纯化后PCR产物直接测序。将测得的序列和推测出的氨基酸序列分别用ClustalX软件与已报道的相应序列比对，用DNASTAR中的MegAlign分析其同源性，并用PAUP绘制系统发育进化树。序列分析结果显示河南猪源隐孢子虫两个分离株Hsp70DNA序列的同源性为99．90．4，与其他隐孢子虫相应序列同源性介于81．9％～99．8％之间，其中与猪隐孢子虫（Cryptosporidiumsuis，AF221533）同源性最高分别为99．8％，97．9％；与安氏隐孢子虫（C．andersoni，AY954592）同源性最低。两个分离株推导Hsp70氨基酸序列的同源性为100％，同其他隐孢子虫相关序列的同源性在92．8％～99．8％之间。本研究为隐孢子虫诊断及流行病学研究打下了良好基础。
To assess the genotype of Cryptosporidium, two Cryptosporidia isolates from pigs in Henan province were obtained. According to Hsp70 gene sequence of Cryptosporidium and other Apicomplexa in GenBank,a pair of Cryptosporidia special primers were designed and synthesized. The fragment of 1948 bp was amplified by PCR from the isolates and then sequenced and compared with the sequences and deduced amino acid sequences with those published for other strains/genus by DNASTAR and Clustal X software before phylogenetic trees were established by the software PAUP. The result showed that the homology between different Cryptosporidium was ranging between 81.9 %～ 99.8. The similarity of the two isolated strains was 99.9%,and homology between the two isolated strains and C. suis（AF221533） were 99.8% and 97. 9%. The lowest similarity was noted between the isolates from pigs and other published Cryptosporidium sequences C. andersoni （AY954592）. The similarity between the two deduced Hsp70 ami;no acid sequence was 100%, and the similarity between the two deduced sequences and other Cryptosporidia sequences ranged from 92.8% to 99.8%. The results indicated that PCR was suitable for the diagnosis and survey of cryptosporidiosis in pigs and provided important information for the diagnosis and epidemic survey of Cryptosporidium infection.
出处
《中国兽医杂志》
CAS
北大核心
2007年第4期6-9,共4页
Chinese Journal of Veterinary Medicine
基金
国家自然科学基金资助项目(30371079)
