Journal of Pure and Applied MicrobiologyVol. 8 No. 6

PCR-RFLP of Staphylococcus aureus Coagulase Gene Isolated from Bovine Subclinical Mastitis

Taruna Bhati1, Prerna Nathawat2, Irfan Ahmad Mir1*, Sandeep Kumar Sharma3, Rahul Yadav1 and Anil Kumar Kataria1

1Department of Veterinary Microbiology and Biotechnology, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Science, Bikaner-334001, India. 2Centre for Studies on Wildlife management and Health, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Science,Bikaner-334001, India. 3Department of Veterinary Microbiology and Biotechnology, Post Graduate Institute of Veterinary Education and Research- Jaipur, Rajasthan University of Veterinary and Animals Sciences, Bikaner (Rajasthan), India.

Received on 06 September 2014 and accepted on 20 November 2014

 

ABSTRACT

The present study was carried out to characterize Staphylococcus aureus isolated from subclinical cases of mastitis in cattle by PCR-RFLP using coagulase (coa) gene. In our study, 38 isolates of S. aureus recovered from Holstein-Friesian (H-F) crossbred and Rathi (a native breed) cattle with subclinical mastitis were subjected to coa gene amplification. Out of 38 isolates, 35 strains produced single amplicon of either 400, 490, 510, 550, 600, 710, 760, 810 or 850 bp while three isolates did not produce any coa amplicon. Subsequently, the products were digested with restriction enzyme AluI for typing of coa gene. Results showed higher coa gene polymorphism of S. aureus isolated from native breed (eight coa types) as compared to that in H-F crossbred cattle (three coa types). The Alu restriction endonuclease generated four and seven RFLP patterns with isolates from H-F crossbred and Rathi cattle respectively. The RFLP patterns obtained from similar amplicons in isolates from these two breeds did not show any variation.

Keywords : Staphylococcus aureus, Mastitis, coa gene, PCR, RFLP.