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

B. Boufana, G. Umhang, J. Qui, X. Chen, S. Lahmar, S. Boue, David Jenkins, P Craig

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Abstract

In order to investigate echinococcosis in co-endemic regions, three 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, E. granulosus G1 and E. 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 3 assays for DNA detection was between 2-10pg. Ethanol precipitation of negative PCR fecal samples was used in order 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.
Original languageEnglish
Pages (from-to)795-802
Number of pages8
JournalAmerican Journal of Tropical Medicine and Hygiene
Volume88
Issue number4
DOIs
Publication statusPublished - Apr 2013

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