A comparison of loop-mediated isothermal amplification (LAMP) with other surveillance tools for Echinococcus granulosus diagnosis in canine definitive hosts

PLoS One. 2014 Jul 9;9(7):e100877. doi: 10.1371/journal.pone.0100877. eCollection 2014.

Abstract

Background: Cystic echinococcosis is highly prevalent in northwest China. A cost-effective, easy to operate diagnostic tool with high sensitivity and specificity would greatly facilitate the monitoring of Echinococcus infections in canine definitive hosts.

Methods: The primers used in the LAMP assay were based on the mitochondrial nad5 gene of E. granulosus sensu stricto (E. granulosus s.s., or E.g.s.s.) and were designed using Primer Explorer V4 software. The developed LAMP assay was compared with a conventional PCR method, copro-ELISA and microscopy, using the faeces of dogs experimentally infected with E.g.s.s., and field-collected faeces of domestic dogs including 190 from Qinghai province highly endemic for E.g.s.s. and 30 controls from an area in Gansu, where a domestic dog de-worming program was in operation.

Results: The positivity rates obtained for the field-collected faecal samples were 12.6%, 1.6% and 2.1% by the LAMP, PCR and copro-ELISA assays, respectively. All samples obtained from the control dogs were negative. Compared with the conventional PCR, the LAMP assay provided 88.8% specificity and 100% sensitivity. The higher sensitivity of the LAMP method was also shown by the fact that it could detect the presence of laboratory challenge dog infections of E. granulsous s.s. four days earlier than the PCR method. Three copro-samples shown positive by the commercial copro-ELISA were all negative by LAMP, PCR and microscopy, which suggests these samples may have originated from another infection rather than E. granulsous s.s., possibly E. shiquicus or E. Canadensis, which is also present in China.

Conclusions: We have developed a potentially useful surveillance tool for determining the prevalence of canine E. granulosus s.s. infections in the field. The LAMP assay may lead to a more cost-effective and practicable way of tracking Echinococcus infections in canids, especially when combined with the copro-ELISA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Helminth / genetics
  • Dog Diseases / diagnosis*
  • Dog Diseases / parasitology
  • Dogs
  • Echinococcosis / diagnosis
  • Echinococcosis / parasitology
  • Echinococcosis / veterinary*
  • Echinococcus granulosus / genetics*
  • Enzyme-Linked Immunosorbent Assay
  • Epidemiological Monitoring
  • Feces / parasitology
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques
  • Prevalence
  • Sensitivity and Specificity

Substances

  • DNA, Helminth

Grants and funding

This study was financially supported by Gansu Provincial Key Science and Technology Projects (1203NKDA039); Special Fund for Agro-scientific Research in the Public Interest (Grant No. 201303037; Grant No. 200903036-07), the People's Republic of China; Natural Science Foundation of China (Grant No. 30960339); and National Health and Medical Research Councils (NHMRCs) Project (APP-1009539), Australia. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.