A multiplex PCR for differential detection of Echinococcus granulosus sensu stricto,Echinococcus multilocularis and Echinococcus canadensis in China  

作　　者：Jing-Ye Shang Guang-Jia Zhang Sha Liao Yan Huang Wen-Jie Yu Wei He Guang-You Yang Tiao-Ying Li Xing-Wang Chen Bo Zhong Qian Wang Qi Wang Rui-Rui Li Hao Wang 

机构地区：[1]Institute of Parasitic Diseases,Sichuan Center for Disease Control and Prevention,Chengdu,Sichuan,People's Republic of China [2]Department of Parasitology,College of Veterinary Medicine,Sichuan Agricultural University,Chengdu,Sichuan,People’s Republic of China

出　　处：《Infectious Diseases of Poverty》2019年第4期49-56,共8页贫困所致传染病（英文）

基　　金：the Scientific Research Project of Health and Family Planning Commission of Sichuan Province(No.17PJ440).

摘　　要：Background:Echinococcosis caused byEchinococcus is one of the most major infectious diseases in north-west highland of China.E.granulosus sensu strict,E.multilocularis,andE.canadensis are known to be the only three species related to human health transmitting in the areas.To achieve targeted treatment and control of echinococcosis,the accurate identification and discrimination of the species are important.However,currently the available diagnostic approaches do not present ideal results either in accuracy or efficiency.Methods:In the study,a set of primers were designed to aim at the three human-pathogenicEchinococcus species in China.The one-step multiplex PCR assay was developed and evaluated for the specificity and sensitivity.A total of 73parasitic lesions and 41 fecal materials obtained from human and various animals collected in the clinic and the field were tested to assess the applicability of this method.Results:The multiplex PCR effectively detected the individual DNA from the targeted species and their random mixtures generating with distinguishable expected size of products.The detection limit of the assay for each of the three species was 5 pg/μl when they were tested separately.When DNA mixtures of the targeted species containing the same concentration were used as templates,the lowest amount of DNA which can be detected was 50 pg/μl,10 pg/μl and 5 pg/μl forE.granulosus s.s.,E.multilocularis,andE.canadensis respectively.No cross-reactivity was observed when DNA from eight genetically close species was used as control templates.The multiplex PCR identifications of all samples were in line with the original sequencing results except for those infected withE.shiquicus,which showed negative signals in the developed assay.Of all the tested stool materials,16 were previously found positive forEchinococcus by visual and microscopic examination.Among these 16 samples,13 were confirmed by the multiplex PCR,and the other three tested negative.Additionally,the multiplex PCR identified another 14 positive f

关 键 词：ECHINOCOCCOSIS Echinococcus granulosus s.s. Echinococcus multilocularis Echinococcus canadensis Multiplex PCR 

分 类 号：R73[医药卫生—肿瘤]