Molecular Detection of Virulence Factors Genes for Staphylococcus aureus in Diabetic Foot Ulcers in Iraq

Main Article Content

Sarah Zghair Hussein
Ghada Mohammed Saleh

Abstract

One of the main complications in patients with Diabetes Mellitus is diabetic foot ulcers (DFUs), a significant and common disease. The wound of a diabetic foot reduces  vascular supply and decrease host immune response, facilitating bacterial infection. The pathogen that is most commonly isolated in DFUs is Staphylococcus aureus. The purpose of this work was to identify S. aureus by conventional, biochemical, and molecular detection of the virulence factor genes in DFUs. This case cohort study was directed at Al-Hussein Teaching Hospital (AIsamawa, Iraq) and Medical City/Baghdad Hospital (Baghdad, Iraq) from January to July 2022, and it involved 140 patients with diabetic foot ulcers. Extraction of bacterial DNA from the swab samples was done by ordinary microbiology techniques and was used to achieve polymerase chain reaction (PCR) using specific gene primers (16S ribosomal RNA, mecA, pvl, and α-hly). S. aureus was identified in 34 (24.28%) of the samples. 73 (52.14%) positive cultures of bacterial growth versus 67 (47.85%) show negative results for culturing. The concentration of DNA extracted from 140 swab samples ranged from 10 to 153 ng/µl. 16S rRNA showed that all 34 isolates gave positive results. PCR results were included. 31 (29.8%) of the Staphylococcus aureus were hiding the mecA gene. The α-hly gene was found in 30 (28.8%) of the S. aureus positive samples. The pvl gene was only present in 10 (9.6%) of S. aureus isolates found in DFU’s samples. In DFUs, S. aureus predominated among the microorganisms. All S. aureus isolates included in this investigation had a high prevalence of the 16S rRNA gene. It serves as a genetic marker that can be utilized to confirm isolates of S. aureus. MecA, pvl, and α-hly genes can be found via PCR analysis.

Article Details

How to Cite
[1]
Hussein, S.Z. and Mohammed Saleh , G. 2024. Molecular Detection of Virulence Factors Genes for Staphylococcus aureus in Diabetic Foot Ulcers in Iraq. Ibn AL-Haitham Journal For Pure and Applied Sciences. 37, 3 (Jul. 2024), 98–105. DOI:https://doi.org/10.30526/37.3.3431.
Section
Biology

How to Cite

[1]
Hussein, S.Z. and Mohammed Saleh , G. 2024. Molecular Detection of Virulence Factors Genes for Staphylococcus aureus in Diabetic Foot Ulcers in Iraq. Ibn AL-Haitham Journal For Pure and Applied Sciences. 37, 3 (Jul. 2024), 98–105. DOI:https://doi.org/10.30526/37.3.3431.

Publication Dates

Received

2023-05-06

Accepted

2023-06-13

Published Online First

2024-07-20

References

American Diabetes Association. Standards of medical care in diabetes—2016 abridged for primary care providers. Clinical diabetes: a publication of the American Diabetes Association. 2016, 34(1), 3.‏ doi: 10.2337/diaclin.34.1.3

Mohammed, S. I.; Mikhael, E. M.; Ahmed, F. T.; Al-Tukmagi, H. F.; Jasim, A. L.. Risk factors for occurrence and recurrence of diabetic foot ulcers among Iraqi diabetic patients. Diabetic foot & ankle 2016, 7(1), 29605.‏ https://doi.org/10.3402/dfa.v7.29605.

Ogba, O.M.; Nsan, E.; Eyam, E.S. Aerobic bacteria associated with diabetic foot ulcers and their susceptibility pattern. Biomedical Dermatology. 2019, 3(1), 1-6.‏ 1 https://doi.org/10.1186/s41702-019-0039-x .

Hamad, S.L.; Melconian, A.K.A.. Staphylococcus aureus Nasal Carriage and Obesity among Patients with Type Two Diabetes Mellitus. Iraqi Journal of Science. 2016, 2840-2848.‏ https://ijs.uobaghdad.edu.iq/index.php/eijs/article/view/6258.

Gholizadeh, P.; Mahallei, M.; Pormohammad, A.; Varshochi, M.; Ganbarov, K.; Zeinalzadeh, E.; Mahmood, S.S.; Kafil, H. S. Microbial balance in the intestinal microbiota and its association with diabetes, obesity and allergic disease. Microbial Pathogenesis. 2019, 127, 48-55.‏ https://doi.org/10.1016/j.micpath.2018.11.031.

hettigar, K.; Murali, T. S. Virulence factors and clonal diversity of Staphylococcus aureus in colonization and wound infection with emphasis on diabetic foot infection. European Journal of Clinical Microbiology & Infectious Diseases. 2020, 39(12), 2235-2246.‏ https://doi.org/10.1007/s10096-020-03984-8.

Kadhim, B. A.; Essa, M. A.; Sabbah, M. R. Prevalence of enterotoxin type a in Staphylococcus aureus isolated from different sources. Iraqi Journal of Agricultural Sciences. 2022, 53(6), 1325-1330.‏ https://doi.org/10.36103/ijas.v53i6.1648.

AlSaadi, B.Q.H.; Mohaisen, S.; Kazaal, Z.; Abdulla, Z. N. Study the prevalence of mecA gene in methicillin resistance Staphylococcus aureus (MRSA) isolated from different clinical specimens and their antibiotic resistance profile. Annals of Tropical Medicine and Public Health. 2020, 23, 231-223.‏ http://doi.org/10.36295/ASRO.2020.231223.

Ismail, M.C.. Relationship between Alpha-toxin production and biofilm for1nation by Staphylococcus aureus isolated from Eye's infections. Ibn AL-Haitham Journal For Pure and Applied Science. 2017 20(1), 13-18.‏ https://jih.uobaghdad.edu.iq/index.php/j/article/view/1307.

Divyakolu, S.; Chikkala, R.; Ratnakar, K. S.; Sritharan, V. Hemolysins of Staphylococcus aureus—An Update on Their Biology, Role in Pathogenesis and as Targets for Anti-Virulence Therapy. Advances in Infectious Diseases. 2019, 9(2), 80-104.‏ DOI: 10.4236/aid.2019.92007.

Alwash, S. J.; Aburesha, R. A. Detection of toxin-associated genes in seven spa-types of Staphylococcus aureus in Iraq. Indian Journal of Forensic Medicine & Toxicology. 2021, 15(3), 4156-4164.‏ DOI: 10.37506/ijfmt.v15i3.15945.

Jauneikaite, E.; Ferguson, T.; Mosavie, M.; Fallowfield, J. L.; Davey, T.; Thorpe, N.; Lamb, L. E. Staphylococcus aureus colonization and acquisition of skin and soft tissue infection among Royal Marines recruits: a prospective cohort study. Clinical microbiology and infection. 2020, 26(3), 381-e1.‏ https://doi.org/10.1016/j.cmi.2019.07.014.

Jafar Alwash, S.; Abed Aburesha, R. The Differences in Antibiotic-resistance among Several Staphylococcus aureus strains in Iraq. Medico-legal Update. 2021, 21(3).‏ DOI: 10.37506/mlu.v21i3.3034.

Abd Zaid, A. M.; Kandala, N. J. Identification of Methicillin Resistant Staphylococcus aureus using touchdown pcr and phenotypic methods from patients and hospitals environments in different Iraqi cities. Iraqi Journal of Agricultural Sciences. 2021, 52(6), 1356-1364.‏ https://doi.org/10.36103/ijas.v52i6.1475.

Ibraheem, H. T.; Al-Mathkhury, H. J. F. pvl-carried methicillin resistant Staphylococcus aureus isolated from hospitalized patients in Baghdad, Iraq: none. Iraqi Journal of Science. 2018, 1967-1972.‏ https://ijs.uobaghdad.edu.iq/index.php/eijs/article/view/468.

Delgado, S.; García, P.; Fernández, L.; Jiménez, E.; Rodríguez-Baños, M.; del Campo, R.; Rodríguez, J. M. Characterization of Staphylococcus aureus strains involved in human and bovine mastitis. FEMS Immunology and Medical Microbiology. 2011, 62(2), 225-235.‏ https://doi.org/10.1111/j.1574-695X.2011.00806.x.

Gumaa, M. A.; Idris, A. B.; Bilal, N. E.; Hassan, M. A. First insights into molecular basis identification of 16s ribosomal RNA gene of Staphylococcus aureus isolated from Sudan. BMC Research Notes. 2021, 14(1), 1-5.‏ https://doi.org/10.1186/s13104-021-05569-w.

Gadban, T. H.; Al-Amara, S. S.; Jasim, H. A. Screening the frequency of panton-valentine leukocidin (pvl) gene between methicillin resistant Staphylococcus aureus isolated from diabetic foot patients in Al-Basrah governorate, south of Iraq. Systematic Reviews in Pharmacy. 2020, 11(11), 285-290.‏ http://faculty.uobasrah.edu.iq/uploads/publications/1614500670.pdf.

Sotto, A.; Richard, J.L.; Messad, N.; Molinari, N.; Jourdan, N.; Schuldiner, S.; French Study Group on the Diabetic Foot. Distinguishing colonization from infection with Staphylococcus aureus in diabetic foot ulcers with miniaturized oligonucleotide arrays: a French multicenter study. Diabetes Care, 2012, 35(3), 617-623.‏ https://doi.org/10.2337/dc11-1352.

Vu, B. G.; Stach, C. S.; Salgado-Pabón, W.; Diekema, D. J.; Gardner, S. E.; Schlievert, P. M. Superantigens of Staphylococcus aureus from patients with diabetic foot ulcers. The Journal of Infectious Diseases. 2014, 210(12), 1920-1927.‏ https://doi.org/10.1093/infdis/jiu350.

Sandhu, S.; Rathnayake, I.; Huygens, F. Prevalence of methicillin resistance and virulence determinants of Staphylococcus aureus in diabetic foot ulcers. International Journal of Basic and Clinical Pharmacology, 2014, 3(6), 978-982.‏ https://doi.org/10.5455/2319-2003.ijbcp20141201.

Al-Hassnawi, H. H.; Al-Charrakh, A. H.; Al-Khafaji, J. K. Occurrence of mecA, SCCmec IV, pvl, lukED Genes in Community Acquired Methicillin Resistance Staphylococcus aureus (CA-MRSA) from Hilla/Iraq. Medical Journal of Babylon, 2013, 10(1).‏ https://www.uobabylon.edu.iq/publications/medicine_edition15/medicine15_16.doc.

Gordon, R. J.; Lowy, F.D. Pathogenesis of methicillin-resistant Staphylococcus aureus infection. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2008, 46 Suppl 5(Suppl 5), S350–S359. https://doi.org/10.1086/533591.

Ayepola, O.O.; Olasupo, N.A., Egwari, L.O.; Schaumburg, F. Characterization of Panton-Valentine leukocidin-positive Staphylococcus aureus from skin and soft tissue infections and wounds in Nigeria: a cross-sectional study. F1000Research. 2018, 7, 1155. https://doi.org/10.12688/f1000research.15484.1.

He, C.; Xu, S.; Zhao, H.; Hu, F.; Xu, X.; Jin, S.; Liu, Q. Leukotoxin and pyrogenic toxin Superantigen gene backgrounds in bloodstream and wound Staphylococcus aureus isolates from eastern region of China. BMC Infectious Diseases, 2018, 18(1), 1-10.‏ https://doi.org/10.1186/s12879-018-3297-0.

Li, Y.; Song, Y.; Huang, Z.; Mei, L.; Jiang, M.; Wang, D.; Wei, Q. Screening of Staphylococcus aureus for Disinfection Evaluation and Transcriptome Analysis of High Tolerance to Chlorine-Containing Disinfectants. Microorganisms. 2023, 11, 475. https://doi.org/10.3390/microorganisms11020475.

Kim, C.; Milheiriço, C.; Gardete, S.; Holmes, M.A.; Holden, M.T.; de Lencastre, H.; Tomasz, A. (Properties of a novel PBP2A protein homolog from Staphylococcus aureus strain LGA251 and its contribution to the β-lactam-resistant phenotype. The Journal of Biological Chemistry. 2012, 287(44), 36854–36863. https://doi.org/10.1074/jbc.M112.395962.

Peacock, S.J.; Paterson, G.K. Mechanisms of Methicillin Resistance in Staphylococcus aureus. Annual Review of Biochemistry. 2015, 84, 577–601. https://doi.org/10.1146/annurev-biochem-060614-034516.

Kumar, S.; Singh, S.; Kumar, V.; Datta, S.; Dhanjal, D. S.; Sharma, P.; Singh, J. Pathogenesis and antibiotic resistance of Staphylococcus aureus. In Model Organisms for Microbial Pathogenesis, Biofilm Formation and Antimicrobial Drug Discovery. 2020, 99-115.‏ https://link.springer.com/chapter/10.1007/978-981-15-1695-5_7.

Raheema, R.H.; Melek, H.K.; Jassim, A.T. Characterization of antibiotic resistance of Staphylococcus aureus isolated from patients with diabetic foot ulcers in Wasit Province. International Journal of Science and Research Archive. 2021, 3(2), 201-208.‏ https://doi.org/10.30574/ijsra.2021.3.2.0146.