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

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 Echi...

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Bibliographic Details
Published in:The American journal of tropical medicine and hygiene 2013-04, Vol.88 (4), p.795-802
Main Authors: Boufana, Belgees, Umhang, Gérald, Qiu, Jiamin, Chen, Xingwang, Lahmar, Samia, Boué, Franck, Jenkins, David, Craig, Philip
Format: Article
Language:English
Subjects:
Animals
China - epidemiology
DNA, Helminth - analysis
DNA, Helminth - genetics
DNA, Mitochondrial - analysis
DNA, Mitochondrial - genetics
Dogs - parasitology
Echinococcosis - epidemiology
Echinococcosis - parasitology
Echinococcosis - veterinary
Echinococcus granulosus
Echinococcus granulosus - classification
Echinococcus granulosus - enzymology
Echinococcus granulosus - genetics
Echinococcus multilocularis
Echinococcus multilocularis - classification
Echinococcus multilocularis - enzymology
Echinococcus multilocularis - genetics
Endemic Diseases
Ethanol - chemistry
Feces - parasitology
Foxes - parasitology
Genes, Mitochondrial
Genotype
Genotyping Techniques - methods
Mitochondria - genetics
NADH Dehydrogenase - genetics
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Sequence Analysis, DNA
Tibet - epidemiology
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Summary: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.
ISSN:0002-9637
1476-1645
DOI:10.4269/ajtmh.12-0331