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Scholars Journal of Applied Medical Sciences | Volume-8 | Issue-03
Amplification Based Detection of Four Target Genes of Staphylococcus Isolates by Multiplex PCR
Dr. M. Mallika, Dr. L. Prashanthi, Dr. N. Padmapriya, Dr. G. Jyothi Lakshmi
Published: March 30, 2020 | 141 89
DOI: 10.36347/sjams.2020.v08i03.048
Pages: 1048-1053
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Abstract
Background: MRSA infections have become a global health problem. The community acquired MRSA has been increasingly reported. Hence characterization of these strains is important to prevent the spread of infections in hospitals and community. Aim: The present study evaluated the amplification based detection of four target genes of Staphylococcus isolates by multiplex PCR. Method: A total of 100 exudate samples from suspected cases of Orthopaedic infections were collected. Majority of the isolates were Staphylococcus, which were identified and speciated based on cultural characteristics and biochemical reactions and coagulase test. Antibiogram was done by Kirby Bauer disc diffusion and Methicillin resistance by cefoxitin disc diffusion method. Amplification based detection of four target genes of Staphylococcus isolates was done by multiplex PCR. Result: Out of total 100 samples processed. 27 Gram positive cocci were isolated. Out of 27, 23 were Staphylococcus aureus,4 were CONS. Out of all staphylococci isolated (27), 20 Staphylococcus aureus and 1 out of 4 CONS showed methicillin resistance .Multiplex PCR confirmed the results as Genus staphylococcus by the presence of 16SrRNA (100%), species by the presence of femA (95.8%), methicillin resistance by the presence of mecA (87.5%) and community acquired MRSA by the presence of luk PVL (40%).Conclusion: Multiplex PCR assay was found to be rapid and accurate procedure for confirmation of genus, species ,methicillin resistance and community acquired strains. Hence multiplex PCR is considered as Gold standard.