Rapid Detection of MRSA by Loop Mediated Isothermal Amplification in Bovine Milk Samples
G. Sathish
Madras Veterinary College, Chennai, Tamilnadu, PIN-600007, India
E. Hemakumar
Chettinad Academy of Research and Education, Chennai, Tamilnadu, PIN-603103, India
Kurunchi C. Divya *
Chettinad Academy of Research and Education, Chennai, Tamilnadu, PIN-603103, India
*Author to whom correspondence should be addressed.
Abstract
Aim: To develop a simple and rapid detection method for diagnosis of MRSA in bovine milk samples suspected for mastitis.
Methodology: The laboratory sensitivity and specificity of LAMP assay was carried out using available laboratory strains. Milk samples were collected from Thiruvallur and Kanchipuram districts of Tamilnadu, India. DNA was directly isolated from the milk samples and mecA gene was screened by both PCR and LAMP methods.
Results: The LAMP assay successfully amplified the mecA gene under isothermal conditions at 64°C within 60 min. LAMP assay was able to detect 10 pg of DNA and did not amplify mecA gene from bacterial DNA of other species. The screening of milk samples for MRSA showed 47 Positive out of 77 Samples by PCR and 43 positive out of 77 Samples by LAMP.
Conclusion: Application of LAMP assay enabled rapid and easy detection of MRSA in milk samples suspected for bovine mastitis.
Keywords: MRSA, bovine mastitis, LAMP assay, milk samples, mecA gene