Document Type : Original Article

Authors

1 Medical Laboratory Technique Department, The Islamic University, Diwaniya, Iraq

2 Research and Studies Department, The Islamic University, Najaf, Iraq

Abstract

Staphylococcus aureus contains numerous surface proteins called microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) that mediate binding to host tissues and initiate colonization leading to infection. Virulence genes such as enzymes, toxins, adhesin proteins, cell surface proteins play an important role in the pathogenicity of S. aureus strains. The distribution and prevalence of virulence genes vary among S. aureus strains in different regions. However, the highest frequency of virulence genes among S. aureus strains is related to toxin genes. There are many PCR methods for detecting these microorganisms such as conventional PCR, multiplex PCR, reverse transcription PCR (RT-PCR), and quantitative PCR (qPCR). Therefore, this study aimed to investigate the presence of virulence genes among methicillin-resistant S. aureus (MRSA) strains. In this study, multiplex PCR technique was applied to determine the presence of virulence genes among MRSA strains. Results showed the frequency of virulence genes among bacterial strains isolated from Al-Najaf Al-Ashraf teaching hospital. In addition, among the strains, hla gene with 91% frequency, exhibited the highest prevalence among pathogenic genes. Sea, mecA, clfB, femA, fnbB, tsst, hlb genes with 88%, 65%, 54%, 45%, 39%, 27% and 13% were in the next ranks, respectively. This investigation showed mecA is a gene found in bacterial cells that allows them to be resistant to antibiotics such as methicillin and other penicillin-like antibiotics. 

Graphical Abstract

Evaluation of virulence factors among Staphylococcus aureus strains isolated from patients with urinary tract infection in Al-Najaf Al-Ashraf teaching hospital

Keywords

Main Subjects

Selected author of this article by journal

ِDr. Qassim hassan Aubais aljelehawy
The Islamic University

Researchgate ; Google Scholar

Open Access

This article is licensed under a CC BY License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit: http://creativecommons.org/licenses/by/4.0/

 

Publisher’s Note

CMBR journal remains neutral with regard to jurisdictional claims in published maps and institutional afflictions.

 

Letters to Editor

Given that CMBR Journal's policy in accepting articles will be strict and will do its best to ensure that in addition to having the highest quality published articles, the published articles should have the least similarity (maximum 15%). Also, all the figures and tables in the article must be original and the copyright permission of images must be prepared by authors. However, some articles may have flaws and have passed the journal filter, which dear authors may find fault with. Therefore, the editor of the journal asks the authors, if they see an error in the published articles of the journal, to email the article information along with the documents to the journal office.

CMBR Journal welcomes letters to the editor ([email protected], [email protected]) for the post-publication discussions and corrections which allows debate post publication on its site, through the Letters to Editor. Critical letters can be sent to the journal editor as soon as the article is online. Following points are to be considering before sending the letters (comments) to the editor.

[1] Letters that include statements of statistics, facts, research, or theories should include appropriate references, although more than three are discouraged.

[2] Letters that are personal attacks on an author rather than thoughtful criticism of the author’s ideas will not be considered for publication.

[3] There is no limit to the number of words in a letter.

[4] Letter writers should include a statement at the beginning of the letter stating that it is being submitted either for publication or not.

[5] Anonymous letters will not be considered.

[6] Letter writers must include Name, Email Address, Affiliation, mobile phone number, and Comments.

[7] Letters will be answered as soon as possible.

  1. Abbas-Al-Khafaji ZK, Aubais-aljelehawy Qh (2021) Evaluation of antibiotic resistance and prevalence of multi-antibiotic resistant genes among Acinetobacter baumannii strains isolated from patients admitted to al-Yarmouk hospital. Cellular, Molecular and Biomedical Reports 1(2):60-68
  2. Alavi M, Rai M (2021) Antisense RNA, the modified CRISPR-Cas9, and metal/metal oxide nanoparticles to inactivate pathogenic bacteria. Cellular, Molecular and Biomedical Reports 1(2):52-59
  3. Almasian-Tehrani N, Alebouyeh M, Armin S, Soleimani N, Azimi L, Shaker-Darabad R (2021) Overview of typing techniques as molecular epidemiology tools for bacterial characterization. Cellular, Molecular and Biomedical Reports 1(2):69-77
  4. Alavi M, Adulrahman NA, Haleem AA, Al-Râwanduzi ADH, Khusro A, Abdelgawad MA, Ghoneim MM, Batiha GE-S, Kahrizi D, Martinez F, Koirala N (2022) Nanoformulations of curcumin and quercetin with silver nanoparticles for inactivation of bacteria. Cellular and Molecular Biology 67(5):151-156. doi:https://doi.org/10.14715/cmb/2021.67.5.21
  5. Aljelehawy Q, Karimi N, Alavi M (2021) Comparison of antibacterial and cytotoxic activities of phytosynthesized ZnONPs by leaves extract of Daphne mucronata at different salt sources. Materials Technology 36(12):747-759. doi:https://doi.org/10.1080/10667857.2020.1794280
  6. Alavi M, Karimi N (2022) Antibacterial, hemoglobin/albumin-interaction, and molecular docking properties of phytogenic AgNPs functionalized by three antibiotics of penicillin, amoxicillin, and tetracycline. Microbial Pathogenesis 164:105427. doi:https://doi.org/10.1016/j.micpath.2022.105427
  7. Becker K, Skov RL, von Eiff C (2015) Staphylococcus, Micrococcus, and Other Catalase-Positive Cocci. In: Manual of Clinical Microbiology. pp 354-382. doi:https://doi.org/10.1128/9781555817381.ch21
  8. Diekema DJ, Pfaller MA, Schmitz FJ, Smayevsky J, Bell J, Jones RN, Beach M (2001) 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. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America 32 Suppl 2:S114-132. doi:https://doi.org/10.1086/320184
  9. Stein GE, Wells EM (2010) The importance of tissue penetration in achieving successful antimicrobial treatment of nosocomial pneumonia and complicated skin and soft-tissue infections caused by methicillin-resistant Staphylococcus aureus: vancomycin and linezolid. Current Medical Research and Opinion 26(3):571-588. doi:https://doi.org/10.1185/03007990903512057
  10. Kadariya J, Smith TC, Thapaliya D (2014) Staphylococcus aureus and Staphylococcal Food-Borne Disease: An Ongoing Challenge in Public Health. BioMed Research International 2014:827965. doi:https://doi.org/10.1155/2014/827965
  11. Tuomanen EI, Que Y-A, François P, Haefliger J-A, Entenza J-M, Vaudaux P, Moreillon P (2001) Reassessing the Role of Staphylococcus aureus Clumping Factor and Fibronectin-Binding Protein by Expression in Lactococcus lactis. Infection and Immunity 69(10):6296-6302. doi:https://doi.org/10.1128/IAI.69.10.6296-6302.2001
  12. Foster TJ (2016) The remarkably multifunctional fibronectin binding proteins of Staphylococcus aureus. European Journal of Clinical Microbiology & Infectious Diseases 35(12):1923-1931. doi:https://doi.org/10.1007/s10096-016-2763-0
  13. Grumann D, Nübel U, Bröker BM (2014) Staphylococcus aureus toxins – Their functions and genetics. Infection, Genetics and Evolution 21:583-592. doi:https://doi.org/10.1016/j.meegid.2013.03.013
  14. Tuomanen EI, Karlsson A, Saravia-Otten P, Tegmark K, Morfeldt E, Arvidson S (2001) Decreased Amounts of Cell Wall-Associated Protein A and Fibronectin-Binding Proteins in Staphylococcus aureus sarA Mutants due to Up-Regulation of Extracellular Proteases. Infection and Immunity 69(8):4742-4748. doi:https://doi.org/10.1128/IAI.69.8.4742-4748.2001
  15. Oliveira D, Borges A, Simões M (2018) Staphylococcus aureus Toxins and Their Molecular Activity in Infectious Diseases. Toxins 10(6):252
  16. Otto M (2010) Staphylococcus colonization of the skin and antimicrobial peptides. Expert Review of Dermatology 5(2):183-195. doi:https://doi.org/10.1586/edm.10.6
  17. Peschel A, Otto M (2013) Phenol-soluble modulins and staphylococcal infection. Nature Reviews Microbiology 11(10):667-673. doi:https://doi.org/10.1038/nrmicro3110
  18. Houri H, Samadpanah M, Tayebi Z, Norouzzadeh R, Malekabad ES, Dadashi A-R (2020) Investigating the toxin profiles and clinically relevant antibiotic resistance genes among Staphylococcus aureus isolates using multiplex-PCR assay in Tehran, Iran. Gene Reports 19:100660. doi:https://doi.org/10.1016/j.genrep.2020.100660
  19. Budzyńska A, Skowron K, Kaczmarek A, Wietlicka-Piszcz M, Gospodarek-Komkowska E (2021) Virulence Factor Genes and Antimicrobial Susceptibility of Staphylococcus aureus Strains Isolated from Blood and Chronic Wounds. Toxins 13(7). doi:https://doi.org/10.3390/toxins13070491
  20. Loges LA, Silva DB, Paulino GVB, Landell MF, Macedo AJ (2020) Polyketides from marine-derived Aspergillus welwitschiae inhibit Staphylococcus aureus virulence factors and potentiate vancomycin antibacterial activity in vivo. Microbial Pathogenesis 143:104066. doi:https://doi.org/10.1016/j.micpath.2020.104066
  21. Arciola CR, Campoccia D, Montanaro L (2018) Implant infections: adhesion, biofilm formation and immune evasion. Nature Reviews Microbiology 16(7):397-409. doi:https://doi.org/10.1038/s41579-018-0019-y
  22. Xiong M, Zhao J, Huang T, Wang W, Wang L, Zhao Z, Li X, Zhou J, Xiao X, Pan Y, Lin J, Li Y (2020) Molecular Characteristics, Virulence Gene and Wall Teichoic Acid Glycosyltransferase Profiles of Staphylococcus aureus: A Multicenter Study in China. Frontiers in Microbiology 11. doi:https://doi.org/10.3389/fmicb.2020.02013
  23. Wang X, Liu Q, Zhang H, Li X, Huang W, Fu Q, Li M (2018) Molecular Characteristics of Community-Associated Staphylococcus aureus Isolates From Pediatric Patients With Bloodstream Infections Between 2012 and 2017 in Shanghai, China. Frontiers in Microbiology 9. doi:https://doi.org/10.3389/fmicb.2018.01211
  24. Zhao H, Xu S, Yang H, He C, Xu X, Hu F, Shu W, Gong F, Zhang C, Liu Q (2019) Molecular Typing and Variations in Amount of tst Gene Expression of TSST-1-Producing Clinical Staphylococcus aureus Isolates. Frontiers in Microbiology 10. doi:https://doi.org/10.3389/fmicb.2019.01388
  25. Zi C, Zeng D, Ling N, Dai J, Xue F, Jiang Y, Li B (2018) An improved assay for rapid detection of viable Staphylococcus aureus cells by incorporating surfactant and PMA treatments in qPCR. BMC Microbiology 18(1):132. doi:https://doi.org/10.1186/s12866-018-1273-x
  26. Fraunholz M, Sinha B (2012) Intracellular Staphylococcus aureus: live-in and let die. Frontiers in cellular and infection microbiology 2:43. doi:https://doi.org/10.3389/fcimb.2012.00043
  27. Mehrotra M, Wang G, Johnson WM (2000) Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1, and methicillin resistance. Journal of clinical microbiology 38(3):1032-1035. doi:https://doi.org/10.1128/JCM.38.3.1032-1035.2000
  28. Eftekhar F, Rezaee R, Azad M, Azimi H, Goudarzi H, Goudarzi M (2017) Distribution of Adhesion and Toxin Genes in Staphylococcus aureus Strains Recovered From Hospitalized Patients Admitted to the ICU. Arch Pediatr Infect Dis 5(1):e39349. doi:https://doi.org/10.5812/pedinfect.39349
  29. Tavares A, Nielsen JB, Boye K, Rohde S, Paulo AC, Westh H, Schønning K, de Lencastre H, Miragaia M (2014) Insights into alpha-hemolysin (Hla) evolution and expression among Staphylococcus aureus clones with hospital and community origin. PLoS One 9(7):e98634-e98634. doi:https://doi.org/10.1371/journal.pone.0098634
  30. Kmieciak W, Szewczyk EM, Ciszewski M (2016) Searching for Beta-Haemolysin hlb Gene in Staphylococcus pseudintermedius with Species-Specific Primers. Curr Microbiol 73(1):148-152. doi:https://doi.org/10.1007/s00284-016-1038-4
  31. Costa AM, Kay I, Palladino S (2005) Rapid detection of mecA and nuc genes in staphylococci by real-time multiplex polymerase chain reaction. Diagnostic microbiology and infectious disease 51(1):13-17. doi:https://doi.org/10.1016/j.diagmicrobio.2004.08.014
  32. Kamarehei F, Ghaemi EA, Dadgar T (2013) Prevalence of enterotoxin A and B genes in Staphylococcus aureus isolated from clinical samples and healthy carriers in Gorgan City, North of Iran. Indian journal of pathology & microbiology 56(3):265-268. doi:https://doi.org/10.4103/0377-4929.120388
  33. Kreiswirth BN, Projan SJ, Schlievert PM, Novick RP (1989) Toxic shock syndrome toxin 1 is encoded by a variable genetic element. Reviews of infectious diseases 11 Suppl 1:S83-88; discussion S88-89. doi:https://doi.org/10.1093/clinids/11.supplement_1.s83
  34. Nagao M, Okamoto A, Yamada K, Hasegawa T, Hasegawa Y, Ohta M (2009) Variations in amount of TSST-1 produced by clinical methicillin resistant Staphylococcus aureus (MRSA) isolates and allelic variation in accessory gene regulator (agr) locus. BMC Microbiol 9:52. doi:https://doi.org/10.1186/1471-2180-9-52
  35. Xie Y, He Y, Gehring A, Hu Y, Li Q, Tu S-I, Shi X (2011) Genotypes and Toxin Gene Profiles of Staphylococcus aureus Clinical Isolates from China. PLoS One 6(12):e28276. doi:https://doi.org/10.1371/journal.pone.0028276
  36. Pakbaz Z, Sahraian MA, Sabzi S, Mahmoodi M, Pourmand MR (2017) Prevalence of sea, seb, sec, sed, and tsst-1 genes of Staphylococcus aureus in nasal carriage and their association with multiple sclerosis. Germs 7(4):171-177. doi:https://doi.org/10.18683/germs.2017.1123
  37. Thalsø-Madsen I, Torrubia FR, Xu L, Petersen A, Jensen C, Frees D (2019) The Sle1 Cell Wall Amidase Is Essential for β-Lactam Resistance in Community-Acquired Methicillin-Resistant Staphylococcus aureus USA300. Antimicrob Agents Chemother 64(1):e01931-01919. doi:https://doi.org/10.1128/AAC.01931-19
  38. Galdiero E, Liguori G, D'Isanto M, Damiano N, Sommese L (2003) Distribution of mecA among methicillin-resistant clinical staphylococcal strains isolated at hospitals in Naples, Italy. European Journal of Epidemiology 18(2):139-145. doi:https://doi.org/10.1023/A:1023067930211
  39. Choi SM, Kim SH, Kim HJ, Lee DG, Choi JH, Yoo JH, Kang JH, Shin WS, Kang MW (2009) Multiplex PCR for the Detection of Genes Encoding Aminoglycoside Modifying Enzymes and Methicillin Resistance among Staphylococcus Species. jkms 18(5):631-636. doi:https://doi.org/10.3346/jkms.2003.18.5.631
  40. Askari E, Soleymani F, Arianpoor A, Tabatabai SM, Amini A, Naderinasab M (2012) Epidemiology of mecA-Methicillin Resistant Staphylococcus aureus (MRSA) in Iran: A Systematic Review and Meta-analysis. Iran J Basic Med Sci 15(5):1010-1019
  41. Li X, Xiong Y, Fan X, Feng P, Tang H, Zhou T (2012) The role of femA regulating gene on methicillin-resistant Staphylococcus aureus clinical isolates. Médecine et Maladies Infectieuses 42(5):218-225. doi:https://doi.org/10.1016/j.medmal.2008.09.031
  42. Kobayashi N, Wu H, Kojima K, Taniguchi K, Urasawa S, Uehara N, Omizu Y, Kishi Y, Yagihashi A, Kurokawa I (1994) Detection of mecA, femA, and femB genes in clinical strains of staphylococci using polymerase chain reaction. Epidemiology and Infection 113(2):259-266. doi:https://doi.org/10.1017/S0950268800051682
  43. Breves A, Miranda CAC, Flores C, Filippis Id, Clementino MM (2015) Methicillin-and vancomycin-resistant Staphylococcus aureus in health care workers and medical devices. Jornal brasileiro de patologia e medicina laboratorial 51:143-152. doi: https://doi.org/10.5935/1676-2444.20150025
  44. Xiao M, Zhao R, Zhang Q, Fan X, O’Sullivan MVN, Li D-F, Wang X-Y, Wu H-L, Kong F, Xu Y-C (2016) Genotypic Diversity of Staphylococcus aureus α-Hemolysin Gene (hla) and Its Association with Clonal Background: Implications for Vaccine Development. PLoS One 11(2):e0149112. doi:https://doi.org/10.1371/journal.pone.0149112
  45. Zhang D-X, Li Y, Yang X-Q, Su H-Y, Wang Q, Zhang Z-H, Liu Y-C, Tian C-L, Cui C-C, Liu M-C (2020) In vitro Antibiotic Susceptibility, Virulence Genes Distribution and Biofilm Production of Staphylococcus aureus Isolates from Bovine Mastitis in the Liaoning Province of China. Infect Drug Resist 13:1365-1375. doi:https://doi.org/10.2147/IDR.S247765
  46. Ren Q, Liao G, Wu Z, Lv J, Chen W (2020) Prevalence and characterization of Staphylococcus aureus isolates from subclinical bovine mastitis in southern Xinjiang, China. Journal of Dairy Science 103(4):3368-3380. doi:https://doi.org/10.3168/jds.2019-17420
  47. Nemati M, Hermans K, Devriese LA, Maes D, Haesebrouck F (2009) Screening of genes encoding adhesion factors and biofilm formation in Staphylococcus aureus isolates from poultry. Avian Pathology 38(6):513-517. doi:https://doi.org/10.1080/03079450903349212
  48. Burke FM, McCormack N, Rindi S, Speziale P, Foster TJ (2010) Fibronectin-binding protein B variation in Staphylococcus aureus. BMC Microbiology 10(1):160. doi:https://doi.org/10.1186/1471-2180-10-160
  49. Kot B, Szweda P, Frankowska-Maciejewska A, Piechota M, Wolska K (2016) Virulence gene profiles in Staphylococcus aureus isolated from cows with subclinical mastitis in eastern Poland. Journal of Dairy Research 83(2):228-235. doi:https://doi.org/10.1017/S002202991600008X