摘要
目的隐孢子虫病是一种人兽共患寄生虫病,在艾滋病病人和儿童中的感染率可分别高达48%和17.5%,可导致严重的水样腹泻甚至危及生命,免疫功能正常者也能感染出现急性自限性腹泻。本文通过不同软件对三株人源隐孢子虫分离株进行种类鉴定。方法参照Xiao L H 1999年发表的资料合成隐孢子虫18S rDNA特异性引物(上游引物:5’-AACCTGGTTGATCCTGCCAGTAGTC -3’,下游引物:5’-TGAGCCTTCTGCAGGTTCACCTACG-3’,由大连宝生物工程技术公司合成。),FCR 扩增,产物测序、序列分别用Paup4.0、rhylip3.6、DNAstar4.0等软件分析并与12株隐孢子虫相应序列进行同源性分析,绘制了系统发育进化树,分析鉴定。1、2号株采自非免疫抑制病人,3号株采自免疫抑制病人。结果①三株隐孢子虫均能扩增出1700bp左右的目的片段。②构建系统发生树;用DNAstar4.0软件进行分析:1与2号株的同源性为99.8%,1和2号株与 AF108864 C.parvum-cattle的同源性分别为99.5%和99.3%,与AF112569 C.hominis进化关系较远。3号株与AF112574 C. meleagridis(火鸡源 C.meleagridis)的同源性为99.8%,3号株与C.suis(AF108861)的同源性为98.9%,而与AF108864 C.parvum -cattle、AF112569 C.hominis、AF108865 C.parvum(Homo sapiens)、AY642591 C.muris、AB210854 C.canis、AF108862 C. felis等均不处于同一进化枝,亲缘关系较远。利用PAUP4.0、Phylip3.6的方法与DNAstar4.0 软件分析的结果基本一致。结论 1和2号分离株均为C.parvum牛基因型,3号分离株是C.meleagridis(火鸡隐孢子虫)。本文结果为深入研究人隐孢子虫病的分子流行病学打下基础。
object: Cryptosporidiosis is paratise of the zoonoses, the rate of infection is respectively 48% and 17. 5% in AIDS and childrens, It can cause heavy watery diarrher and even threaten life. immunocompetent individuals can also infect and occur acute diarrhea, the topic is to identified the three species of cryptosporidium by using different software.. Methods: to synthesize 18S rDNA specific primer by referring to datum published in 1999 by xiao L H, ( upstream primers :5'- AACCTGGTTGATCCTGCCAGTAGTC - 3, downstream primers : 5 ? TGAGCCTTCTGCAGGTTCACCTACG? 3,primer was synthesized by biology enginer technology company. ), PCR amplify, production sequenced. The measured sequence was analysed by the software of Paup4.0 ,Phylip3.6 ,DNAstar4.0 and analysed homologily of 12 cryptosporidium corresponding sequence . and protract the cladogram of phyiogenesis. results:①the three cryptosporidium can amplify aimed segment of about 1700bp. ②construct the cladogram of phylogenesis was analysed by the software of DNAstar4.0 : the homology of 1# 2# is 99.8% , the homology of 1# 2# AF108864 C. parvum-cattle is respectively 99.5%, 99.3% ,and relation of evolution of AF112569 C, hominis is very far. the homology of 3# and AF112574 C. meleagridis is 99.8% , the homology of 3# C. suis is 98.9% ,and AF108864 C. parvum - cattle、AF112569 C. hominis 、AF108865 C. parvum ( Homo sapiens) 、AY642591 C. muris、 AB210854 C. canis 、AF108862 C. fells is located differently tree of evolution, the relation is very far. the result analysed by using the method of PAUP4.0 、Phvlip3.6 and the software of DNAstar4.0 are consistent . conclusion : 1# 2# are C. parvum, 3# is C. meleagridis, the result of the article lay good foundations for molecule epidemiological study of cryptosporidiosis.
出处
《河南预防医学杂志》
2006年第2期79-79,共1页
Henan Journal of Preventive Medicine