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Comparison of AS-PCR, CRS-PCR and PCR-PIRA methods used for screening CVM carriers in Holstein cattle
Kozet Avanus*, Ahmet Altinel
Department of Animal Breeding and Husbandry, Faculty of Veterinary Medicine, Istanbul University, Avcilar, Istanbul 34320, Turkey
Correspondence to: Tel: +90-530-349-9915; Fax: +90-212-473-7241; E-mail: avanus@istanbul.edu.tr
Received: March 31, 2017; Revised: June 6, 2017; Accepted: July 26, 2017; Published online: September 20, 2017.
Abstract
Complex vertebral malformation (CVM) is an inherited and autosomal recessive disorder of Holstein cattle. The aim of this study was comparing sensitivity, specificity, positive and negative predictive values, accuracy and rapidity of three methods; allele specific PCR (AS-PCR), created restriction site PCR (CRS-PCR) and PCR with primer introduced restriction analysis (PCR-PIRA) that were used in identification of CVM carriers in Holstein cattle population. In order to screen G>T mutation in solute carrier family 35 member A3 (SLC35A3) gene DNA sequencing was used as gold standard method. The prevalence of carriers and mutant allele frequency was found 3.2% and 0.016 respectively among Holstein cattle in Thrace region. Among three methods the fastest but the least accurate one was detected as AS-PCR. Although rapidity of CRS-PCR and PCR-PIRA methods was nearly equal, accuracy of PCR-PIRA was found higher than CRS-PCR method. Therefore among three methods PCR-PIRA method might be suggested to use for screening of mutant allele in order to identify CVM carriers in Holstein cattle population.
Keywords: Holstein, cattle, CVM, carrier, Turkey


© 2017 The Korean Society of Veterinary Science.