广东省主要并殖吸虫DNA序列分析  

Identification of Paragonimus species in Guangdong province of China with DNA sequence analysis

作　　者：陈少华[1] 陆予云[1] 朱福祺 李旭文 刘建军[1] 黄志斌[1] 蓝境文 饶鸿[1] 林李丽 谭宁[1] 

机构地区：[1]广州医科大学卫生职业技术学院,广东广州510925 [2]江西卫生职业学院,江西南昌330201 [3]从化市中医医院,广东广州510925

出　　处：《热带医学杂志》2017年第3期337-342,F0003,共7页Journal of Tropical Medicine

基　　金：广东省医学科研基金(A2014620);广州市教育局科研基金(10A198);广州医学院卫生职业技术学院重点项目(2012005)

摘　　要：目的确定广东省主要并殖吸虫虫种。方法采用形态学特征观察和分子生物学DNA测序相结合的研究方法,鉴定广东省主要并殖吸虫虫种。采集卫氏并殖吸虫、三平正并殖吸虫和斯氏并殖吸虫自然疫源地山溪中溪蟹并检获三种并殖吸虫囊蚴,分别人工感染家犬或家猫,饲养60~120 d后,剖杀检获成虫分别制成玻片大体标本。结果三种不同并殖吸虫疫源地蟹体检获的囊蚴鉴定为卫氏并殖吸虫、三平正并殖吸虫和斯氏并殖吸虫。三种吸虫成虫样本的COI基因与ITS2基因进行PCR,扩增产物进行测序并分别与Gen Bank中检索的AY618799.1号基因、AF159594.1号基因、AY140693.1号基因、AY618733.1号基因、AF159602.1号基因和AB713404.1号基因序列比对,同源性分别在99%~100%之间。结论广东省主要并殖吸虫为卫氏并殖吸虫、斯氏并殖吸虫和三平正并殖吸虫。三种并殖吸虫的COI基因与ITS2基因序列与Gen Bank中检获的基因序列同源性分别在99%~100%之间,三种并殖吸虫与Gen Bank中检获虫种无明显差异性。Objective To identify the predominant Paragonimus species in Guangdong province. Methods The major species of Paragonimus in Guangdong province that cause human infections based on morphological and DNA sequence data were identified. Results Domestic dogs and cats were used as experimentally definitive hosts. Three types of Paragonimus metacercariaes were obtained from fresh water crabs in an endemic area, where P.westermani, Euparagonimus cenocopiosis, and Pagumogonimus skrjabini had previously been reported. Subsequently, the metacereariaes were fed to domestic dogs and cats. An autopsy was made on the dogs and cats 60-120 days after infections and revealed adult worms. Gross inspection of the worms suggested that the specimens were P. westermani, Euparagonimus cenocopiosis, and Pagumogonimus skrjabini. The COl and ITS2 gene fragments of three adult worms were amplified with polymerase chain reaction （PCR） and amplified products were used to perform sequence analysis. The amplified products were sequenced and compared with AY618799.1, AF159594.1, AYI40693.I, AY618733.1, AF159602.1, and AB713404.1 in GenBank database. The comparison displayed a similarity range of 99%-100%. Conclusion The major Paragonimus species that are human pathogens in Guangdong province were P. westermani, Pagumogonimus skrjabini, and Euparagonimus cenocopiosis. The COI and ITS2 sequences of the three types of worms showed high degrees of similarity to those in the GenBank （99%-100%）, without significant differences.

关 键 词：并殖吸虫 囊蚴 DNA序列 广东 

分 类 号：R383.23[医药卫生—医学寄生虫学]