A critical assessment of two real-time PCR assays targeting the (SSU) rRNA and gdh genes for the molecular identification of Giardia intestinalis in a clinical laboratory

J Clin Pathol. 2014 Sep;67(9):811-6. doi: 10.1136/jclinpath-2014-202224.

Abstract

Introduction: Giardiasis is an intestinal diarrhoeal illness caused by the flagellate protozoan parasite Giardia intestinalis. Molecular techniques for the identification of G. intestinalis have generally been shown to offer a better detection rate of the parasite than the traditional faecal concentration and microscopy techniques.

Aim: The aim of this study was to critically assess the performance of a commercial and a published real-time PCR assay for their potential use as frontline tests for the diagnosis of giardiasis.

Methods: A composite reference standard of enzyme immunoassay and rapid membrane test was used in a diagnostic accuracy study to assess the performance of Primerdesign's, and Verweij et al G. intestinalis real-time PCR assays, comparing them with the traditional ova, cysts and parasite microscopy test (OCP-M).

Results: The Verweij real-time PCR used primers for the (SSU) rRNA gene, and produced a diagnostic sensitivity of 93.4% (95% CI 88.30% to 98.50%) and an efficiency of 100%. Primerdesign's real-time PCR used primers for the glutamate dehydrogenase gene and produced a diagnostic sensitivity of 61.5% (95% CI 51.50% to 71.50%) and an efficiency of 203%. The OCP-M sensitivity was 83.5% (95% CI 75.87% to 91.13%).

Conclusions: The Verweij real-time PCR was robust and the most sensitive assay suited for use as a first-line diagnostic test for giardiasis.

Publication types

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

MeSH terms

  • Calibration
  • DNA, Protozoan / genetics*
  • Feces / parasitology
  • Giardia lamblia / classification
  • Giardia lamblia / genetics*
  • Giardia lamblia / isolation & purification
  • Glutamate Dehydrogenase / genetics*
  • Humans
  • Laboratories, Hospital*
  • Microscopy
  • Predictive Value of Tests
  • Protozoan Proteins / genetics*
  • RNA, Ribosomal / genetics*
  • Real-Time Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction / standards
  • Reference Standards
  • Reproducibility of Results

Substances

  • DNA, Protozoan
  • Protozoan Proteins
  • RNA, Ribosomal
  • Glutamate Dehydrogenase