A Clinical Report on Molecular Identification of Cryptosporidium sp. by nested PCR

Abstract

The intra cellular protozoan parasite Cryptosporidium sp. is one of the major causes of diarrhea in calf and other animals. During our study period we collected 50 diarrheic faeces samples of calves from 3 different districts (Chittagong, Pabna and Syrajgong) in the country. For diagnosis of Cryptosporidium sp. we have used molecular DNA based tools including highly sensitive nested polymerase chain reaction (nested PCR) and gel electrophoresis to identify the Cryptosporidium sp. from diarrheic faeces of calf below 6 months of age. For nested PCR analysis, specific primers for amplification of 581 base pair fragments of 18s rRNA gene were used. We could not find any positive band in first time reaction, based on this result it can’t sure that all the Templates were negative for DNA, because another investigator found 10% positive result from same sample by Z-N staining and due to time limitation we could not run pcr for several times up to standardization.