IDENTIFICATION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) BY A RAPID PCR TECHNIQUE

Authors

  • Sajjad Mohsin Irayyif
  • R.senthil Kumar
  • Sudhakar Malla

Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important causes of hospital infections worldwide. High-level resistance to methicillin is caused by the mecA gene, which encodes an alternative penicillin-binding protein, PBP2a.  Staphylococcus aureus is an important cause of bloodstream infections and a leading cause of severe health care–associated infections. Laboratory diagnostics S. aureus is a Gram positive, catalase positive aerobic or anaerobic coccus showing hemolytic and large yellow colonies. Staphylococcus aureus strains resistant to methicillin and many other antibiotics are major causes of nosocomial infections worldwide. It is of great necessity to check for those genes and check for the potential to clone partial or major fragments into vectors, which can be a potential candidate for DNA vaccines. Novel methods are being developed for the production of antibodies to specific antigens and thus helping in the process of development of protein based vaccines.

mecA (Methicillin resistant) gene was isolated and ligated into pTZ57R/T cloning vector. The ligated product was then cloned into DH5α strain and allowed to propagate. The plasmids thus cloned were purified and later expressed for the gene of interest in an expression vector. The proteins specific to the gene of interest was then isolated and purified. This proteins purified can in turn be used for protein based vaccines.

Key words: Staphylococcus, MRSA, mecA, Cloning.

Downloads

Download data is not yet available.

References

AK SK, Shetty PJ, Y LS, Chidambaram A, Ranganathan R. Detection of mecA genes of methicillin-resistant Staphylococcus aureus by polymerase chain reaction. Int J Health Rehabil Sci 2012;1(2):64-8.

Anthony BF, Hill HR. Gram-positive bacteria: An overview and summary of session. Rev Infect Dis 1988;10 (Suppl 2):S345-50.

Beck WD, Berger-Bächi B, Kayser FH. Additional DNA in methicillin resistant staphylococcus aureus and molecular cloning of mec-specific DNA. J Bacteriol 1986;165(2):373-8.

Berger-Bächi B, Strässle A, Gustafson JE, Kayser FH. Mapping and characterization of multiple chromosomal factors involved in methicillin resistance in Staphylococcus aureus. Antimicrob Agents Chemother 1992;36(7):1367-73.

Brown DF, Reynolds PE. Intrinsic resistance to beta-lactam antibiotics in Staphylococcus aureus. FEBS Lett 1980;122(2):275-8.

Chambers HF, Sachdeva M, Kennedy S. Binding affinity for penicillin-binding protein 2a correlates with in vivo activity of beta-lactam antibiotics against methicillin-resistant Staphylococcus aureus. J Infect Dis 1990;162(3):705-10.

Diekema DJ, Pfaller MA, Schmitz FJ, Smayevsky J, Bell J, Jones RN, et al. Survey of infections due to Staphylococcus species: Frequency of occurrence and antimicrobial susceptibility of isolates collected in the United States, Canada, Latin America, Europe, and the Western Pacific region for the SENTRY Antimicrobial Surveillance Program, 1997-1999. Clin Infect Dis 2001;32 Suppl 2:S114-32.

Dinges MM, Orwin PM, Schlievert PM. Exotoxins of Staphylococcus aureus. Clin Microbiol Rev 2000;13(1):16-34.

Fitzgerald JR, Sturdevant DE, Mackie SM, Gill SR, Musser JM. Evolutionary genomics of Staphylococcus aureus: Insights into the origin of methicillin-resistant strains and the toxic shock syndrome epidemic. Proc Natl Acad Sci U S A 2001 17;98:8821-6.

Hackbarth CJ, Chambers HF. blaI and blaR1 regulate beta-lactamase and PBP 2a production in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 1993;37(5):1144-9.

Ito T, Hiramatsu K. Acquisition of methicillin resistance and progression of multiantibiotic resistance in methicillin-resistant Staphylococcus aureus. Yonsei Med J 1998;39(6):526-33.

Ito T, Katayama Y, Asada K, Mori N, Tsutsumimoto K, Tiensasitorn C, et al. Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2001;45(5):1323-36.

Kreiswirth B, Kornblum J, Arbeit RD, Eisner W, Maslow JN, McGeer A, et al. Evidence for a clonal origin of methicillin resistance in Staphylococcus aureus. Science 1993;259(5092):227-30.

Kreiswirth B, Kornblum J, Arbeit RD, Eisner W, Maslow JN, McGeer A, et al. Evidence for a clonal origin of methicillin resistance in Staphylococcus aureus. Science 1993;259(5092):227-30.

Lowy FD. Staphylococcus aureus infections. N Engl J Med 1998;339(8):520-32.

Moreillon P, Entenza JM, Francioli P, McDevitt D, Foster TJ, François P, et al. Role of Staphylococcus aureus coagulase and clumping factor in pathogenesis of experimental endocarditis. Infect Immun 1995;63(12):4738-43.

Musser JM, Kapur V. Clonal analysis of methicillin-resistant staphylococcus aureus strains from intercontinental sources: Association of the mec gene with divergent phylogenetic lineages implies dissemination by horizontal transfer and recombination. J Clin Microbiol 1992;30(2):2058-63.

Oliveira DC, Tomasz A, de Lencastre H. Secrets of success of a human pathogen: Molecular evolution of pandemic clones of meticillin-resistant staphylococcus aureus. Lancet Infect Dis 2002;2(2):180-9.

Proctor RA, van Langevelde P, Kristjansson M, Maslow JN, Arbeit RD. Persistent and relapsing infections associated with small-colony variants of Staphylococcus aureus. Clin Infect Dis 1995;20(1):95-102.

Shands KN, Schmid GP, Dan BB, Blum D, Guidotti RJ, Hargrett NT, et al. Toxic-shock syndrome in menstruating women: Association with tampon use and Staphylococcus aureus and clinical features in 52 cases. N Engl J Med 1980;303(25):1436-42.

Uhlén M, Guss B, Nilsson B, Gatenbeck S, Philipson L, Lindberg M. Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications. J Biol Chem 1984;259(3):1695-702.

Wielders CL, Vriens MR, Brisse S, de Graaf-Miltenburg LA, Troelstra A, Fleer A, et al. In-vivo transfer of mecA DNA to Staphylococcus aureus [corrected]. Lancet 2001;357(9269):1674-5.

Published

01-11-2014

How to Cite

Irayyif, S. M., R. Kumar, and S. Malla. “IDENTIFICATION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) BY A RAPID PCR TECHNIQUE”. Asian Journal of Pharmaceutical and Clinical Research, vol. 7, no. 5, Nov. 2014, pp. 14-19, https://mail.innovareacademics.in/journals/index.php/ajpcr/article/view/1695.

Issue

Section

Original Article(s)

Most read articles by the same author(s)