Molecular detection of Clostridium perfringens type D alpha and epsilon toxin genes from various tissues in lambs
Azam Ali Nasir1,3, Muhammad Younus2,*,
Muti-ur-Rehman3, Muhammad Latif3, Asif Rashid1,
Rashid Ahmad1 and Muhammad Abbas1
1Veterinary
Research Institute, Zarrar Shaheed Road, Lahore Cantt-54810; 2Department
of Pathobiology, College of Veterinary and Animal Sciences, Jhang
(sub-campus of UVAS, Lahore); 3Department of Pathology,
University of Veterinary and Animal Sciences, Lahore, Pakistan *Corresponding author: younusrana@hotmail.com
Abstract
Different tissues including duodenal scrapings,
liver, lungs and kidneys of each infected (n=6) and control (n=4) lambs were
subjected to PCR for the identification of alpha and epsilon toxin genes of
Clostridium perfringens type D. Alpha
and epsilon toxin genes were amplified at annealing temperature 52.2 and 50.2oC
with amplicon size about 247bp and 665bp, respectively. A high percentage of
alpha toxin genes of C. perfringens
type D were amplified from duodenal scrapings followed by liver, kidneys lungs
in cultured broth while a low percentage was observed when PCR was attempted
directly on these tissues without prior culturing in media. It was concluded
that distribution of alpha and epsilon
toxin genes varies in different organs being the highest in duodenal tissue and
PCR is suitable assay for the typing of C.
perfringens toxin genes.