the first study of rt-pcr as the identification technique of the cause of bovine viral diarrhea (bvd) in endometrial tissue of cows in iran

Bovine viral diarrhea (bvd) is one of the most important among the viral diseases that cause lots of disturbances in the reproduction system of dairy cattle. possible diagnosis methods are included of traditional virus isolation (vi), serological methods (elisa, ifa and ihc) and molecular methods (rt-pcr, real-time pcr, ddpcr, lamp, rpa and crispr-cas system). rt-pcr is a highly sensitive and specific method which has not been used to diagnose this disease in iran. the purpose of this study was to investigate the prevalence rate of bvd in referred dairy cows to mashhad industrial animal slaughterhouse through the rt-pcr method. uterine endometrial tissue was sampled from the uterus of 33seemingly healthy slaughtered cows in 1.5 ml micro tubes including of rna preservation solution. rna extraction and reverse transcription were done. the pcr time and temperature program included of primary denaturation at 95˚c for 5 minutes and then 40 cycles at 95 ˚c, 61 ˚c, 72 ˚c for 30, 20 and 30 seconds and finally 72 ˚c for 5 minutes respectively. at last, the reaction products were electrophoresed on a 1% agarose gel to determine the test results. one positive sample was observed among 33 tested samples which showed a 3% of prevalence. the outbreak of bvd in dairy cows in iran have been conducted in several studies through non molecular methods with higher or lower degree of prevalence than the present study. the importance of subject is that it is the first study to investigate the spread of bvd on uterine samples of dairy cows in iran by using rt-pcr method. since the abortion by bvd in cows mainly depends on infection of the uterine tissue, perhaps diagnosis of bvd prevalence in the endometrial tissue of the uterus through rt-pcr can provide a more accurate abortion diagnosis for bvd.