Use of 5-Enolpyruvylshikmate-3-Phosphate Synthase Encoding Gene for Typing of Staphylococcus aureus Isolated from Skin and Urinary Tract Infections of Human

Document Type : Original Article

Authors

1 Department of Microbiology, Faculty of Veterinary Medicine, Urmia University,Urmia, Iran

2 Department of Pathology, Faculty of Medicine, University of Medical Sciences, Urmia, Iran

Abstract

Objective(s)
Staphylococcus aureus is both a successful human commensal and a major pathogen. In this study we investigated the genetic diversity of 26 S. aureus isolates recovered from human skin and urinary tract infections.
Materials and Methods
Typing procedure for the studied S. aureus isolates was performed based on PCR amplification of the aroA gene, which encodes the enzyme 5-enolpyruvylshikmate-3-phosphate synthase (EPSPS) that involves in aromatic amino acid biosynthesis, and restriction fragment length polymorphism (RFLP) analysis of the product.
Results
All S. aureus isolates produced a single PCR amplification product of 1,153 bp. Digestion of the PCR products with the TaqI endonuclease revealed four different aroA gene patterns designated as A, B, N and H according to the nomenclature system of previous studies. In general, 80.77% of the studied isolates displayed type N, 7.69% were type B, 7.69% were type H and 3.85% displayed type A.
Conclusion
Divergent aroA types were detected among S. aureus isolates from skin and urinary tract infections. The results showed that urinary tract infections were contaminated by S. aureus isolates with identical banding patterns (A), while isolates recovered from skin infections had different aroA types. This study also indicates that aroA genotypes vary not only from region to region, but also in individual hosts within a region.

Keywords

References

1. Gould IM. Antibiotics, skin and soft tissue infection and meticillin-resistant Staphylococcus aureus: Cause and effect. Int J Antimicrob Agents 2009; 34:S8-11.
2. McCaig LF, McDonald LC, Mandal S, Jernigan DB. Staphylococcus aureus-associated skin and soft tissue infections in ambulatory care. Emerg Infect Dis 2006; 12:1715-1723.
3. Bowler PG, Duerden BI, Armstrong DG. Wound microbiology and associated approaches to wound management. Clin Microbiol Rev 2001; 14:244-269.
4. Charalambous CP, Zipitis CS, Kumar R, Lipsett PA, Hirst P, Paul A. Soft tissue infections of the extremities in an orthopedic centre in the UK. J Infect 2003; 46:106-110.
5. Mohanty S, Kapil A, Dhawan B, Das BK. Bacteriological and antimicrobial susceptibility profile of soft tissue infections from Northern India. Indian J Med Sci  2004; 58:10-15.
6. Verma S, Joshi S, Chitnis V, Hemwani N, Chitnis D. Growing problem of methicillin resistant Staphylococci-Indian scenario. Indian J Med Sci  2000; 54:535-540.
7. Fluit AC, Schmitz FJ, Verhoef J. Frequency of isolation of pathogens from bloodstream, nosocomial pneumonia, skin and soft tissue, and urinary tract infections occurring in European patients. Eur J Clin Microbiol Infect Dis 2001; 20:188-191.
8. Ong PY, Ohtake T, Brandt C, Strickland I, Boguniewicz M, Ganz T, et al. Endogenous antimicrobial peptides and skin infections in atopic dermatitis. N Engl J Med 2002; 347:1151-1160.
9. Niyonsaba F, Ogawa H. Protective roles of the skin against infection: Implication of naturally occurring human antimicrobial agents beta-defensins, cathelicidin ll-37 and lysozyme. J Dermatol Sci 2005; 40:157-168.
10.  Josefsson E, Kubica M, Mydel P, Potempa J, Tarkowski A. In vivo sortase a and clumping factor A mRNA expression during Staphylococcus aureus infection. Microb Pathog 2008; 44:103-110.
11. van Leeuwen WB, Melles DC, Alaidan A, Al-Ahdal M, Boelens HA, Snijders SV, et al. Host- and tissue-specific pathogenic traits of Staphylococcus aureus. J Bacteriol 2005; 187:4584-4591.
12. Gadepalli R, Dhawan B, Kapil A, Sreenivas V, Jais M, Gaind R, et al. Clinical and molecular characteristics of nosocomial meticillin-resistant Staphylococcus aureus skin and soft tissue isolates from three Indian hospitals. J Hosp Infect 2009; 73:253-263.
13.  Laplana LM, Cepero MA, Ruiz J, Zolezzi PC, Calvo MA, Erazo MC, et al. Molecular typing of Staphylococcus aureus clinical isolates by pulsed-field gel electrophoresis, Staphylococcal cassette chromosome mec type determination and dissemination of antibiotic resistance genes. Int J Antimicrob Agents 2007; 30:505-513.
14.  Montesinos I, Salido E, Delgado T, Cuervo M, Sierra A. Epidemiologic genotyping of methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis at a University hospital and comparison with antibiotyping and protein a and coagulase gene polymorphisms. J Clin Microbiol 2002; 40:2119-2125.
15.  Hookey JV, Edwards V, Cookson BD, Richardson JF. PCR-RFLP analysis of the coagulase gene of Staphylococcus aureus: Application to the differentiation of epidemic and sporadic methicillin-resistant strains. J Hosp Infect 1999; 42:205-212.
16.  Wichelhaus TA, Hunfeld KP, Boddinghaus B, Kraiczy P, Schafer V, Brade V. Rapid molecular typing of methicillin-resistant Staphylococcus aureus by PCR-RFLP. Infect Control Hosp Epidemiol 2001; 22:294-298.
17.  Casey AL, Worthington T, Caddick JM, Hilton AC, Lambert PA, Elliott TS. RAPD for the typing of Staphylococcus aureus implicated in nosocomial infection. J Hosp Infect 2007; 66:192-193.
18.  Enright MC, Day NP, Davies CE, Peacock SJ, Spratt BG. Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. J Clin Microbiol 2000; 38:1008-1015.
19.  Strommenger B, Braulke C, Heuck D, Schmidt C, Pasemann B, Nubel U, et al. Spa typing of Staphylococcus aureus as a frontline tool in epidemiological typing. J Clin Microbiol 2008; 46:574-581.
20.  Marcos JY, Soriano AC, Salazar MS, Moral CH, Ramos SS, Smeltzer MS, et al. Rapid identification and typing of Staphylococcus aureus by PCR-restriction fragment length polymorphism analysis of the aroA gene. J Clin Microbiol 1999; 37:570-574.
21.  Fitzgibbon JE, Braymer HD. Cloning of a gene from Pseudomonas sp. strain pg2982 conferring increased glyphosate resistance. Appl Environ Microbiol 1990; 56:3382-3388.
22.  Saei HD, Ahmadi M, Mardani K, Batavani RA. Genotyping of Staphylococcus aureus isolated from bovine mastitis based on PCR-RFLP analysis of the aroA gene. Comp Clin Pathol 2010; 19:163-168.
23.  El-Huneidi W, Bdour S, Mahasneh A. Detection of enterotoxin genes seg, seh, sei, and sej and of a novel aroA genotype in Jordanian clinical isolates of Staphylococcus aureus. Diagn Microbiol Infect Dis 2006; 56:127-132.
24.  Brakstad OG, Aasbakk K, Maeland JA. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J Clin Microbiol 1992; 30:1654-1660.
25.  Kim CH, Khan M, Morin DE, Hurley WL, Tripathy DN, Kehrli M, Jr, et al. Optimization of the PCR for detection of Staphylococcus aureus nuc gene in bovine milk. J Dairy Sci 2001; 84:74-83.
26.  Chmelnitsky I, Navon-Venezia S, Leavit A, Somekh E, Regev-Yochai G, Chowers M, et al. SCCmec and spa types of methicillin-resistant Staphylococcus aureus strains in Israel. Detection of SCCmec type V. Eur J Clin Microbiol Infect Dis  2008; 27:385-390.
27.  Fenner L, Widmer AF, Dangel M, Frei R. Distribution of spa types among methicillin-resistant Staphylococcus aureus isolates during a 6 year period at a low-prevalence University hospital. J Med Microbiol  2008; 57:612-616.
28.  Deurenberg RH, Vink C, Kalenic S, Friedrich AW, Bruggeman CA, Stobberingh EE. The molecular evolution of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect 2007; 13:222-235.
29.  Vandenesch F, Naimi T, Enright MC, Lina G, Nimmo GR, Heffernan H, et al. Community-acquired methicillin-resistant Staphylococcus aureus carrying Panton-Valentine leukocidin genes: Worldwide emergence. Emerg Infect Dis 2003; 9:978-984.
30.  Yamasaki O, Tristan A, Yamaguchi T, Sugai M, Lina G, Bes M, et al. Distribution of the exfoliative toxin D gene in clinical Staphylococcus aureus isolates in France. Clin Microbiol Infect 2006; 12:585-588.
31.  Ruppitsch W, Indra A, Stoger A, Mayer B, Stadlbauer S, Wewalka G, et al. Classifying spa types in complexes improves interpretation of typing results for methicillin-resistant Staphylococcus aureus. J Clin Microbiol 2006; 44:2442-2448.
32.  Vindel A, Cuevas O, Cercenado E, Marcos C, Bautista V, Castellares C, et al. Methicillin-resistant Staphylococcus aureus in Spain: Molecular epidemiology and utility of different typing methods. J Clin Microbiol 2009; 47:1620-1627.
33. Wu D, Wang Q, Yang Y, Geng W, Wang Q, Yu S, et al. Epidemiology and molecular characteristics of community-associated methicillin-resistant and methicillin-susceptible Staphylococcus aureus from skin/soft tissue infections in a children's hospital in Beijing, China. Diagn Microbiol Infect Dis  2010; 67:1-8.
34.  Herron-Olson L, Fitzgerald JR, Musser JM, Kapur V. Molecular correlates of host specialization in Staphylococcus aureus. PLoS One 2007; 2:e1120.
35.  Leitner G, Krifucks O, Glickman A, Younis A, Saran A. Staphylococcus aureus strains isolated from bovine mastitis: Virulence, antibody production and protection from challenge in a mouse model. FEMS Immunol Med Microbiol 2003; 35:99-106.
36.  Kerro Dego O, van Dijk JE, Nederbragt H. Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review. Vet Q 2002; 24:181-198.
37.  Rainard P, Corrales JC, Barrio MB, Cochard T, Poutrel B. Leucotoxic activities of Staphylococcus aureus strains isolated from cows, ewes, and goats with mastitis: Importance of lukm/lukf'-pv leukotoxin. Clin Diagn Lab Immunol  2003; 10:272-277.
38.  Reinoso E, Bettera S, Frigerio C, DiRenzo M, Calzolari A, Bogni C. RAPD-PCR analysis of Staphylococcus aureus strains isolated from bovine and human hosts. Microbiol Res 2004; 159:245-255.
39.  Kapur V, Sischo WM, Greer RS, Whittam TS, Musser JM. Molecular population genetic analysis of Staphylococcus aureus recovered from cows. J Clin Microbiol 1995; 33:376-380.
40.  Nashev D, Toshkova K, Salasia SI, Hassan AA, Lammler C, Zschock M. Distribution of virulence genes of Staphylococcus aureus isolated from stable nasal carriers. FEMS Microbiol Lett 2004; 233:45-52.
41.  Clarke SR, Mohamed R, Bian L, Routh AF, Kokai-Kun JF, Mond JJ, et al. The Staphylococcus aureus surface protein IsdA mediates resistance to innate defenses of human skin. Cell Host Microbe 2007; 1:199-212.
42.  Toshkova K, Annemuller C, Akineden O, Lammler C. The significance of nasal carriage of Staphylococcus aureus as risk factor for human skin infections. FEMS Microbiol Lett 2001; 202:17-24.
43.  Fritz SA, Epplin EK, Garbutt J, Storch GA. Skin infection in children colonized with community-associated methicillin-resistant Staphylococcus aureus. J Infect 2009; 59:394-401. Epub 2009 Sep 9.
44.  Tulloch LG. Nasal carriage in Staphylococcal skin infections. Br Med J 1954; 2:912-913.
45.  Ellis MW, Hospenthal DR, Dooley DP, Gray PJ, Murray CK. Natural history of community-acquired methicillin-resistant Staphylococcus aureus colonization and infection in soldiers. Clin Infect Dis  2004; 39:971-979.
46.  Baba-Moussa L, Anani L, Scheftel JM, Couturier M, Riegel P, Haikou N, et al. Virulence factors produced by strains of Staphylococcus aureus isolated from urinary tract infections. J Hosp Infect  2008; 68:32-38.
47.  Shupp Byrne DE, Sedor JF, Soroush M, McCue PA, Mulholland SG. Interaction of bladder glycoprotein gp51 with uropathogenic bacteria. J Urol  2001; 165:1342-1346.
48.  Karahan ZC, Tekeli A, Adaleti R, Koyuncu E, Dolapci I, Akan OA. Investigation of Panton-Valentine leukocidin genes and SCCmec types in clinical Staphylococcus aureus isolates from Turkey. Microb Drug Resist  2008; 14:203-210.
49.  Tenover FC, Arbeit R, Archer G, Biddle J, Byrne S, Goering R, et al. Comparison of traditional and molecular methods of typing isolates of Staphylococcus aureus. J Clin Microbiol 1994; 32:407-415.
 

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