内科Issue(2)：128-131,123,5.
虫卵ITS-2序列分析鉴别华支睾吸虫和扇棘单睾吸虫的研究
Study on identification of Clonorchis sinensis and Haplorchis taichui by analysis of Egg ITS-2 sequence
摘要
Abstract
Objective To establish an identification method basis on molecular biology techniques for Clonorchis sinensis and Haplorchis taichui.Methods Adult flukes of C.sinensis and H.taichui were collected from Clonorchiasis endemic areas and the eggs were separated from the worms under stereomicroscope.Roundworm,Whipworm,Hookworm and Pinworm were served as control species,all kinds of eggs were divided into five experimental groups according to the species and the number of eggs.DNA from each group was extracted and amplified the sequence of ITS-2,and amplified products were sequenced and homology analyzed ensure that the products were the target fragments.The species of insect were deter-mined according to the electrophoresis of PCR products.Results The electrophoresis of PCR products of C.sinensis and H. taichui display a distinct band at around 400bp,500bp,respectively;electropherogram in PCR products of mixture DNA of two species eggs showed two obvious bands at around 400bp and 500bp.The two nucleotide sequencing which had se-quenced were compared to Blast,show that both of the sequences are highly homologous with C.sinensis and H.taichi in GenBank ,respectively.The PCR products with four kinds of Heligmosomoides polygyrus eggs have no other bands except the objective strips.Conclusion This method can be used to identify C.sinensis and H.taichui,and the results are not af-fected by four kinds of Heligmosomoides polygyrus eggs ,is an identification method for Clonorchis sinensis and Haplorchis taichui with high sensitivity and specificity.关键词
华支睾吸虫/扇棘单睾吸虫/ITS-2/鉴别方法Key words
Clonorchis sinensis/Haplorchis taichui/ITS-2分类
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吕国丽,杨益超,蒋智华,韦海艳,张伟尉,唐雯茜,林源,黄福明,区方奇..虫卵ITS-2序列分析鉴别华支睾吸虫和扇棘单睾吸虫的研究[J].内科,2014,(2):128-131,123,5.基金项目
广西壮族自治区科技厅科学研究与技术开发计划项目(桂科攻0719006-1-2)。 （桂科攻0719006-1-2）
