Abstract

A SYBR® Green based quantitative real-time PCR (qPCR) technique for detection and quantification of Eimeria species during its in vivo developmental phase in the intestinal tissue of broilers was standardized. Sixty intestinal tissue samples were collected from suspected, ailing and dead birds in commercial broiler farms in Tamil Nadu and Andhra Pradesh. All the intestinal samples were screened by a generic ITS-1 based conventional PCR out of which 29 (48.3 percent) were found positive for Eimeria species. A qPCR assay was standardized for detection of genus Eimeria in the intestinal tissue using the 5S ribosomal RNA as the target of amplification. The parasite genome numbers in tissue were estimated and comparisons drawn between generic Eimeria counts and species-specific E. tenella counts indicated presence of species other than E. tenella in caecal tissue. Quantitative real-time PCR was found to be a sensitive quantitative technique to detect Eimeria infection in broilers during the pre-patent and subclinical phase of infection in commercial broilers when compared to genus specific conventional PCR.
 

Keywords : Eimeria, qPCR, absolute quantification