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  • 1 Department of Biology, Rasht Branch, Islamic Azad University, Iran
  • 2 Young Researchers and Elites Club, Lahijan Branch, Islamic Azad University, Iran
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Panton–Valentine leukocidin (pvl) toxin is an important virulence factor of Staphylococcus aureus. The main genes are coa and spa for distinguishing and typing of S. aureus isolates. The aim of this study was to investigate antibiotic resistance, presence of mecA and pvl genes, as well as epidemiological typing of these isolates according to polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method in clinical sample isolated from Rasht city, Iran. A total of 250 clinical samples have been isolated from different hospitals. First, isolates of S. aureus were identified through microbiological methods and their antibiotic sensitivity was determined by disk diffusion agar based on a standard method of Clinical and Laboratory Standards Institute. DNA was extracted by boiling and presence of pvl and mecA genes was investigated by PCR using specific primers. To type these isolates, amplification of fragments of coa and spa genes was done and restriction enzyme digestion pattern was determined by PCR-RFLP method. Among the 250 samples, 50 isolates belonged to S. aureus and results of antibiotic sensitivity showed that 68% (34 samples) of isolates were methicillin resistant. Frequency of mecA and pvl genes among S. aureus isolates were 60% (30 samples) and 20% (10 samples). The PCR of coa gene showed three patterns whereas that of spa gene showed two patterns for enzyme digestion. Result of PCR-RFLP using HaeIII enzymes for coa gene and Bsp1431 for spa gene showed three patterns for enzyme digestion. Recent studies indicated increase in the resistance of S. aureus to different antibiotics, which is a serious problem in the treatment of infections resulting from S. aureus in this region. The result of PCR of pvl showed high frequency of this gene in this region, and coa and spa typing by PCR-RFLP was a useful tool for typing of S. aureus isolates.

  • 1.

    Masalha, M., Borovok, I., Schreiber, R., Aharonowitz, Y., Cohen, G.: Analysis of transcription of the Staphylococcus aureus aerobic class Ib and anaerobic class III ribonucleotide reductase genes in response to oxygen. J Bacteriol 183, 72607272 (2001).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2.

    Tarai, B., Das, P., Kumar, D.: Recurrent challenges for clinicians: Emergence of methicillin-resistant, vancomycin resistance, and current treatment options. J Lab Physicians 5, 7178 (2013).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 3.

    Shorr, A. F.: Epidemiology of Staphylococcal resistance. Clin Infect Dis 45, 171176 (2007).

  • 4.

    Lina, G., Piemont, Y., Godail-Gamot, F., Bes, M., Peter, M. O., Gauduchon, V.: Involvement of Panton–Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis 29, 11281132 (1999).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 5.

    Supersac, G., Prevost, G., Piemont, Y.: Sequencing of leucocidin R from Staphylococcus aureus P83 suggests that Staphylococcal leucocidins and gamma-hemolysin are members of a single, two-component family of toxins. Infect Immun 61, 580587 (1993).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 6.

    Clark, J.: A brief review of Panton–Valentine leukocidin producing staphylococcal infections in the intensive therapy unit. Curr Anaesth Crit Care 19, 330332 (2008).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 7.

    Colin, D. A., Mazurier, I., Sire, S., Finck-Barbancon, V.: Interaction of the two components of leukocidin from Staphylococcus aureus with human polymorphonuclear leukocyte membranes: Sequential binding and subsequent activation. Infect Immun 62, 31843190 (1994).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8.

    Brown, M. L., O’Hara, F. P., Close, N. M., Mera, R. M., Miller, L. A., Suaya, J. A.: Prevalence and sequence variation of Panton–Valentine leukocidin in methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains in the United States. J Clin Microbiol 50, 8690 (2012).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9.

    Simpson, K. H., Bowden, G., Hook, M., Anvari, B.: Measurement of adhesive forces between individual Staphylococcus aureus MSCRAMMs and protein-coated surfaces by use of optical tweezers. J Bacteriol 185, 20312035 (2003).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Shakeri, F., Shojai, A., Golalipour, M., Alang, S. R., Vaez, H., Ghaemi, E. A.: Spa diversity among MRSA and MSSA strains of Staphylococcus aureus in north of Iran. Int J Microbiol 2010, Article ID 351397 (2010).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Ravathi, G., Puri, J., Jain, B. K.: Bacteriology of burns. Burns 24, 347349 (1998).

  • 12.

    Lu, S. Y., Chang, F. Y., Cheng, C. C., Lee, K. D., Huang, Y. C.: Methicillin-resistant Staphylococcus aureus nasal colonization among adult patients visiting emergency department in a Medical Center in Taiwan. PLoS One 6, e18620 (2011).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    Clinical and Laboratory Standards Institute: Performance Standards for Antimicrobial Susceptibility Testing, Vol. 17. Clinical and Laboratory Standards Institute, Wayne, PA, 2007.

    • Search Google Scholar
    • Export Citation
  • 14.

    Ghaznavi-Rad, E., Nor Shamsudin, M., Sekawi, Z., Belkum, A., Neela, V.: A simplified multiplex PCR assay for fast and easy discrimination of globally distributed staphylococcal cassette chromosome mec types in meticillin-resistant Staphylococcus aureus. Med Microbiol 59, 11351139 (2010).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15.

    Moghadam, S., Havaei, A.: Prevalence of methicillin-resistant Staphylococcus aureus carrying Panton–Valentine leukocidin gene in cutaneous infections in the City of Isfahan. J Med Bacteriol 19, 916 (2012).

    • Search Google Scholar
    • Export Citation
  • 16.

    Mostafa, S.: Molecular typing of methicillin resistant Staphylococcus aureus by spa gene polymorphism. Afr J Microbiol Res 7, 755759 (2013).

    • Search Google Scholar
    • Export Citation
  • 17.

    Khosravi, A. D., Hoveizavi, H., Farshadzadeh, Z.: The prevalence of genes encoding leukocidins in Staphylococcus aureus strains resistant and sensitive to methicillin isolated from burn patients in Taleghani Hospital, Ahvaz, Iran. Burns 38, 247251 (2012).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    Zamani, A., Sadeghian, S., Ghaderkhani, J., Alikhani, M. Y., Najafimosleh, M., Taghi Goodarzi, M.: Detection of methicillin-resistance (mec-A) gene in Staphylococcus aureus strains by PCR and determination of antibiotic susceptibility. Ann Microbiol 57, 273276 (2007).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 19.

    Moradi, N., Javadpour, S., Karmostaji, A.: Reduced sensitivity of Staphylococcus aureus to vancomycin. Hormozgan Uni Med Sci 15, 169177 (2011).

    • Search Google Scholar
    • Export Citation
  • 20.

    Lepsanovic, Z., Jeremic, L. P., Lazic, S., Cirkovic, I.: High prevalence and resistance patterns of community-associated methicillin-resistant Staphylococcus aureus in the Pomoravlje Region, Serbia. Acta Microbiol Immunol Hung 63, 8392 (2016).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 21.

    Haghgoo, S., Moaddab, S., Rafi, A.: Study of antibiotic resistance pattern of Staphylococcus aureus strains isolated from blood cultures in Tabriz Shahid Madani Hospital. J Jundishapur 3, 383390 (2012).

    • Search Google Scholar
    • Export Citation
  • 22.

    Karasartova, D., Cavusoglu, Z. B., Turegun, B., Ozsan, M. T., Şahin, F.: Identification of virulence genes carried by bacteriophages obtained from clinically isolated methicillin-resistant Staphylococcus aureus. Acta Microbiol Immunol Hung 63, 433447 (2016).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23.

    Molla-abbaszadeh, H., Mobayen, H., Mirzaei, H.: Identification of Panton–Valentine leukocidin (pvl) genes in Staphylococcus aureus isolated from in-patients of Emam Reza and Shohada Hospitals of Tabriz by real-time PCR. Iran J Med Microbiol 6, 7280 (2013).

    • Search Google Scholar
    • Export Citation
  • 24.

    Otokunefor, K., Sloan, T., Kearns, A. M., James, R.: Molecular characterization and Panton–Valentine leukocidin typing of community-acquired methicillin-sensitive Staphylococcus aureus clinical isolates. J Clin Microbiol 50, 30693072 (2012).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25.

    Cupane, L., Pugacova, N., Berzina, D., Cauce, V., Gardovska, D., Miklasevics, E.: Patients with Panton–Valentine leukocidin positive Staphylococcus aureus infections run an increased risk of longer hospitalisation. Int J Mol Epidemiol Genet 3, 4855 (2012).

    • Search Google Scholar
    • Export Citation
  • 26.

    Osman, N. A. M., Alrayah, I. E., Mohamed, Y. M., El-Eragi, A. M., Eldirdery, M. M., Salih, M. A.: Molecular study of Panton–Valentine leukocidin genes among Staphylococcus aureus clinical isolates in Khartoum State, Sudan. Am J Microbiol Res 3, 107111 (2015).

    • Search Google Scholar
    • Export Citation
  • 27.

    Lima, D. F., Brazao, N. B., Folescu, T. W., Neves, F. P., Ferreira, A. G., Santos, E. A., Marques, E. A., Leao, R. S.: Panton–Valentine leukocidin (PVL) gene carriage among Staphylococcus aureus strains with SCCmec types I, III, IV, and V recovered from cystic fibrosis pediatric patients in Brazil. Diagn Microbiol Infect Dis 78, 5962 (2014).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28.

    Himabindu, M., Muthamilselvan, D. S., Bishi, D. K., Verma, R. S.: Molecular analysis of coagulase gene polymorphism in clinical isolates of methicilin resistant Staphylococcus aureus by restriction fragment length polymorphism based genotyping. Am J Infect Dis 5, 163169 (2009).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 29.

    Lawrence, C., Cosseron, M., Mimoz, O.: Use of coagulase gene typing method for detection of carrier of methicillin resistant Staphylococcus aureus. J Antimicrob Chemother 37, 687696 (1996).

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30.

    Karahan, M., Nuri, M., Cetinkaya, B.: Investigation of virulence genes by PCR in Stapylococcus aureus isolates originated from subclinical bovine mastitis in Turkey. Pak Vet J 31, 249253 (2011).

    • Search Google Scholar
    • Export Citation
  • 31.

    Schmitz, F., Steiert, M., Tichy, H. V.: Typing of methicillin-resistant Staphylococcus aureus isolates from Dusseldorf by six genotypic methods. J Med Microbiol 47, 341351 (1998).

    • Crossref
    • Search Google Scholar
    • Export Citation

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