Correlation between diarrhea severity and oocyst count via quantitative PCR or fluorescence microscopy in experimental cryptosporidiosis in calves

Am J Trop Med Hyg. 2015 Jan;92(1):45-9. doi: 10.4269/ajtmh.14-0488. Epub 2014 Nov 4.

Abstract

Cryptosporidium is an important diarrhea-associated pathogen, however the correlation between parasite burden and diarrhea severity remains unclear. We studied this relationship in 10 experimentally infected calves using immunofluorescence microscopy and real-time polymerase chain reaction (qPCR) (N = 124 fecal samples). The qPCR data were corrected for extraction/amplification efficiency and gene copy number to generate parasite counts. The qPCR and microscopic oocyst quantities exhibited significant correlation (R(2) = 0.33, P < 0.05), however qPCR had increased sensitivity. Upon comparison with diarrhea severity scores (from 0 to 3), a PCR-based count of ≥ 2.6 × 10(5) parasites or an immunofluorescence microscopy count of ≥ 4.5 × 10(4) oocysts were discriminatory predictors of moderate-to-severe diarrhea (versus no-to-mild diarrhea), with accuracies and predictive values of 72-82%. In summary, a quantitative approach for Cryptosporidium can refine predictive power for diarrhea and appears useful for distinguishing clinical cryptosporidiosis versus subclinical infection.

MeSH terms

  • Animals
  • Cattle
  • Cattle Diseases / parasitology*
  • Cattle Diseases / physiopathology
  • Cryptosporidiosis / parasitology*
  • Cryptosporidiosis / physiopathology
  • Diarrhea / parasitology
  • Diarrhea / physiopathology
  • Diarrhea / veterinary*
  • Microscopy, Fluorescence / methods*
  • Oocysts
  • Parasite Egg Count*
  • Polymerase Chain Reaction / methods*