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- W3030005005 abstract "Objective To compare 3 methods(hemi-nested PCR,multiplex PCR and PCR-RFLP),and optimize the method for discrimination of Echinococcus spp.in animals.Methods Hemi-nested PCR primers were designed according to the mitochondria DNA sequence of the Genus Echinococcus with DNAStar software.Fifty-nine samples included E.multilocularis,E.granulosus and Taenia taeniaeformis were collected from Shiqu County.All the samples were amplified with 3 PCR methods.The specificity and sensitivity of 3 methods were also compared.Furthermore,the sensitivity of 3 methods was evaluated.Results All the 59 samples had positive results by the hemi-nested PCR amplification,with the length of the amplified sequence of 561 bp,but the method could not differentiate specific species without sequencing.Meanwhile,both multiplex PCR and PCR-RFLP could discriminate E. multilocularis,E.granulosus,but the false negative rate(47.5%,28/59)was too high to accept.The two methods also amplified 5(62.5%)of E. granulosus,23(47.9%)of E. multilocularis and 3(100%) of Taenia taeniaeformis,respectively.PCR-RFLP method amplified fragments of the same size in E. granulosus G1 strain and Taenia spp.,and thus it could not be used to discriminate them.The sensitivity of hemi-nested PCR was as 1 000 times and 100 times hish as PCR-RFLP and multiplex PCR,respectively.The minimum concentration of E. granulosus DNA detected by hemi-nested PCR was 253 pg/μl.Statistical analysis showed that detection rate of hemi-nested PCR for Echinococcus spp.was significanfly higher than that of PCR-RFLP and multiplex PCR.Conclusion The results suggested that combined use of these 3 methods could enhance the sensitivity of detection of echinococcosis,and facilitate the diagnosis of Echinococcus spp..Key words: Echinococcus; Polymerase chain reaction; Differentiation of species" @default.
- W3030005005 created "2020-06-05" @default.
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- W3030005005 date "2011-05-28" @default.
- W3030005005 modified "2023-09-23" @default.
- W3030005005 title "Comparison of 3 PCR methods for detection of Echinococcus spp.in animals" @default.
- W3030005005 doi "https://doi.org/10.3760/cma.j.issn.1673-4122.2011.03.005" @default.
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