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

Am J Trop Med Hyg. 2013 Apr;88(4):795-802. doi: 10.4269/ajtmh.12-0331. Epub 2013 Feb 25.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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 / classification*
  • Echinococcus granulosus / enzymology
  • Echinococcus granulosus / genetics
  • 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

Substances

  • DNA, Helminth
  • DNA, Mitochondrial
  • Ethanol
  • NADH Dehydrogenase