title: Development of three PCR assays for the differentiation between Echinococcus shiquicus, E. granulosus (G1 genotype), and E. multilocularis DNA in the co-endemic region of Qinghai-Tibet plateau, China.
abstract: To investigate echinococcosis in co-endemic regions, three polymerase chain reaction (PCR) assays based on the amplification of a fragment within the NADH dehydrogenase subunit 1 (ND1) mitochondrial gene were optimized for the detection of Echinococcus shiquicus, Echinococcus granulosus G1, and Echinococcus multilocularis DNA derived from parasite tissue or canid fecal samples. Specificity using parasite tissue-derived DNA was found to be 100% except for E. shiquicus primers that faintly detected E. equinus DNA. Sensitivity of the three assays for DNA detection was between 2 and 10 pg. Ethanol precipitation of negative PCR fecal samples was used to eliminate false negatives and served to increase sensitivity as exemplified by an increase in detection from 0% to 89% of E. shiquicus coproDNA using necropsy-positive fox samples.
authors: Cestode Zoonoses Research Group, School of Environment and Life Sciences, University of Salford, Salford, Greater Manchester, United Kingdom. B.Boufana@salford.ac.uk
authors: Belgees
authors: B
authors: Boufana
authors: Gérald
authors: G
authors: Umhang
authors: Jiamin
authors: J
authors: Qiu
authors: Xingwang
authors: X
authors: Chen
authors: Samia
authors: S
authors: Lahmar
authors: Franck
authors: F
authors: Boué
authors: David
authors: D
authors: Jenkins
authors: Philip
authors: P
authors: Craig
mesh_terms: D000818:Animals; D002681:China / Q000453:epidemiology; D018509:DNA, Helminth / Q000032:analysis* / Q000235:genetics; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics; D004285:Dogs / Q000469:parasitology; D004443:Echinococcosis / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary; D048209:Echinococcus granulosus / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D048210:Echinococcus multilocularis / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D019353:Endemic Diseases; D000431:Ethanol / Q000737:chemistry; D005243:Feces / Q000469:parasitology; D005589:Foxes / Q000469:parasitology; D050259:Genes, Mitochondrial; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods; D008928:Mitochondria / Q000235:genetics; D009245:NADH Dehydrogenase / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D018609:Tibet / Q000453:epidemiology
chemical_list: D018509:DNA, Helminth; D004272:DNA, Mitochondrial; D000431:Ethanol; D009245:NADH Dehydrogenase