Staphylococcus aureus typing by digestion of protein A coding gene using Bsp143I

Abstract

Background: Protein A is the virulence factors of Staphylococcus aureus rolling in its pathogenesis, and its gene is used for typing. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with different enzymes has been used for this action. Objectives: In this study, we used Bsp143I enzyme for digestion of the gene, coding protein A (spa gene) in S. aureus. The bacteria were isolated from patients and healthy carriers in Gorgan, north of Iran. Patients and Methods: DNAs of 128 S. aureus subjects (53 from healthy carriers and 75 from patients) were extracted and amplified using specific primers of the spa gene. The product was digested by Bsp143I enzyme and its pattern was assessed by gel electrophoresis. Results: There were seven spa types among the tested S. aureus samples, among which six types differed in the repeated X region of the spa gene, but the seventh type had a deletion on one of BSP143I restriction sites. The frequency of spa types among isolated S. aureus samples as well as healthy carriers was six and five, respectively. S. aureus isolated from wounds showed the most diverse spa types (five) among clinical samples. Types 1, 2 and 4 were observed in all clinical samples, while only one case of type 3 was identified among patients, whereas this type constituted over 32% of the isolates among carriers. We found seven and four spa types among methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates, respectively. Conclusions: Our results showed that typing the spa gene using PCR-RFLP using Bsp143I was an acceptable method for typing S. aureus. Furthermore, this survey showed that the types in healthy carriers and MSSA were more variable than patient and MRSA isolates, respectively. We used the Bsp143I enzyme, which was not used in any previous studies on the spa gene. The results of this study suggested that we can use PCR-RFLP of spa gene by Bsp143I for molecular typing and sequencing of S. aureus, instead of relatively expensive methods. This method is relatively rapid and inexpensive, and can be accomplished in centers with conventional molecular facilities. © 2014, Ahvaz Jundishapur University of Medical Sciences