A new spa type t5696 MRSA isolated from veterinary medicine student at Universiti Putra Malaysia

Methicillin-resistant Staphylococcus aureus (MRSA) has been spreading worldwide since its
emergence. Swab samples were collected from the nostrils and oral cavity of veterinary medicine
student at Universiti Putra Malaysia. Phenotypic identification of MRSA was made by
employing broth enrichment, catalase and cogulase tests, mannitol fermentation test on mannitol
salt agar (MSA), slide latex agglutination test, growth on selective media, Oxacillin-resistant
screening agar base (ORSAB). Intense blue colonies on ORSAB presumptively confirmed
MRSA. Antibiotic sensitivity tests using 15 antibiotics revealed that the isolate was resistant
against Amikacin 30 μg (AK30), Amoxicillin 25 μg (AML25), Methicillin 10 μg (MET10),
Oxacillin 1 μg (OX1) Streptomycin 10 μg (S10), and Tetracycline 10 μg (TE10) while being
intermediately resistant Oxacillin (OX1) and susceptible to other antibiotics tested. The
minimum inhibitory concentration (MIC) of the isolate was 8 μg/mL. However, detection of the
resistance gene mecA was proved negative. This discrepancy between phenotypic and mecA
detection methods is attributed to the fact that there are other non-mecA dependent Oxacillin
resistance factors in S. aureus. Amplification of the polymorphic X region of the staphylococcal
protein A gene was done by PCR. Purified PCR product was sequenced, aligned and repeat
numbers were assigned. Repeat sequence pattern was assigned a unique spa type, t5696. The
finding hints the importance of further studies by focusing on the occurrence of MRSA in
veterinary professionals and students with effective molecular typing techniques such as spa
typing. This will in turn helps in understanding the similarities and uniqueness of local isolates in
reference to MRSA isolates reported anywhere in the world