Invastigation of 16S rRNA, mecA and nuc genes in coagulase-positive and negative Staphylococci by Real-Time PCR
Abstract
Staphylococcus aureus is a Gram-positive, round-shaped bacterium that is a member of the Firmicutes. It is often positive for catalase and nitrate reduction. Pathogenic strains often promote infections by producing virulence factors such as potent protein toxins, and the expression of a cell-surface protein that binds and inactivates antibodies. Sepsis-related to methicillin-resistant S. aureus (MRSA) has significant morbidity and high mortality rates (15-30%). The methicillin resistance for S. aureus is coded with the mecA gene, while the methicillin sensitivity is coded with the nuc gene, and they are chromosomal. Similarly, it is coded with the coagulase gene for S. aureus (Coa). In this study, the 16S rRNA gene identification by Real-Time PCR was investigated in forty S. aureus isolates, which were cultured at different times in terms of MIC and SIR tests. The isolates used in the study were determined at the gene level in terms of their differences in methicillin resistance gene (MecA), methicillin susceptibility gene (Nuc), coagulase gene (Coa) and in terms of intra-species differences were examined. As a result of the study, Staphylococcus spp. yielded positive results with 16S rRNA gene-specific primers in all isolates. Real-Time PCR analysis of the isolates with SYBRGreen-based PCR analysis was performed with 16S rRNA gene-specific primers, and the samples were confirmed to be Staphylococcus. Analysis at the family level was followed by Coa, Nuc, and MecA gene Real-Time PCR results, and it was found that, in terms of Coa and Nuc genes, 19 isolates were positive and 21 isolates were negative. In terms of MecA gene, 16 isolates were positive according to the positive sigmoidal curves and to the single peak melting values, whereas 24 isolates were found to be negative. It was thought that this study will benefit the community by contributing to the rapid and effective diagnosis and treatment of infections caused by coagulase positive and negative Staphylococci.
References
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- Ashraf A, Hanan AF, Fatma I, Abdelhalim A. Detection of species-specific gene thermonuclease (Nuc) in Staphylococcus aureus from chickens by Real-Time PCR assay. BVMJ. 2014; 27(2): 415‐422.
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Abstract
References
- Altinok I, Balta F, Capkin E, Kayis S. Disease of rainbow trout caused by Pseudomonas luteola. Aquaculture. 2007; 273(4): 393-397.
- Ashraf A, Hanan AF, Fatma I, Abdelhalim A. Detection of species-specific gene thermonuclease (Nuc) in Staphylococcus aureus from chickens by Real-Time PCR assay. BVMJ. 2014; 27(2): 415‐422.
- Bilgehan H. Clinical Microbiological Diagnosis, Methods of Examining Microbiological Morphology. Barış Publications, 2002.
- Elhassan MM, Ozbak HA, Hemeg HA, Elmekki MA, Ahmed LM. Absence of the MecA gene in methicillin resistant Staphylococcus aureus isolated from different clinical specimens in shendi city, Sudan. Biomed Res Int, 2015; 1(1): 1-5.
- Hamidi RM, Hosseinzadeh S, Shekarforoush SS, Poormontaseri M, Derakhshandeh A. Association between the enterotoxin production and presence of Coa, Nuc genes among Staphylococcus aureus isolated from various sources, in Shiraz. Iran J Vet Res, 2015; 16(4): 381.
- Hoegh SV, Skov MN, Boye K, Worning P, Jensen TG, Kemp M. Variations in the Staphylococcus aureus-specific Nuc gene can potentially lead to misidentification of meticillin-susceptible and-resistant S. aureus. J Med Microbiol, 2014; 63(7): 1020-1022.
- Kaya E, Kırmalı O, Doğan Ö, Berk D, Kaya D. The susceptibility of ampicillin and ampicillin-sulbactam to bacteria with chronic otitis media. Türk Mikrobiyol Cem Derg, 2003; 33(1): 115-117.
- Levi K, Towner K. Detection of Methicillin-Resistant Staphylococcus aureus (MRSA) in blood with the EVIGENE MRSA Detection Kit. J Clin Microbiol, 2003; 41(8): 3890-3892.
- Levinson W. Medical Microbiology & Immunology, Özgünen, T., Trans. Ankara: Güneş Bookstore, 2002.
- Nasution GS, Suryanto D, Kusumawati RL. Detection of meca gene from methicillin resistant Staphylococcus aureus isolates of north sumatera. InIOP Conference Series: Environ. Earth Sci, 2018; 130(1): 1-7.
- Nia KM, Sepehri G, Khatmi H, Shakibaie MR. Isolation and antimicrobial susceptibility of bacteria from chronic suppurative otitis media patients in Kerman, Iran. Iran Red Crescent Med J, 2011; 13(12): 891-894.
- Önalan Ş. Expression differences of stress and immunity genes in rainbow trout (Oncorhynchus mykiss, Walbaum 1792) with different bacterial fish diseases. Sr J Aquacult-Bamid, 2019; 1537(1): 1-10.
- Önalan Ş, Yavuz Hİ. Spectrophotometric determination of development differences of Lactococcus garvieae isolates in liquid media containing nanoparticles. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi, 2019; 1(1): 6-15.
- Özen NS, Dağlar D, Baysan BÖ, Yıldırım Ç, Yazısız H, Öğünç D, et al. Evaluation of MRSA id chromogenic media in detecting methicillin resistant Staphylococcus aureus strains. Ankem Derg, 2011; 25(1): 31-34.
- Seidel C, Peters S, Eschbach E, Feßler AT, Oberheitmann B, Schwarz S. Development of a nucleic acid lateral flow immunoassay (NALFIA) for reliable, simple and rapid detection of the methicillin resistance genes MecA and MecC. Vet Microbiol, 2017; 200(1): 101-106.
- Tok D, Coşkun Ö. Antibiotic susceptibilities of microorganisms growing in the cultures of patients with chronic otitis media. TAF Prev Med Bull, 2010; 9(1): 51-54.
- Uçan N. Determination of coagulase negative staphylococci and goat antibiotic resistance in goats with subclinical mastitis. (Master Thesis). Aydın: Adnan Menderes University, Institute of Health Sciences, 2014.
- Willke A, Sayan M, Meriç M, Mutlu M. Early diagnosis of methicillin resistance in Real-Time PCR in Staphylococci growing in blood cultures. Mikrobiyol Bul, 2012; 46(4): 671-675.
- Yadav R, Kumar A, Singh V, Singh J, Yadav SK. Molecular determination of methicillin resistance MecA and virulence Coa genes in Staphylococcus aureus from pyogenic clinical cases of companion animals in India. Turk J Vet Anim Sci, 2018; 42(5): 371-375.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Journal Section | 2022 Volume 6 Number 1 |
| Authors | |
| Early Pub Date | March 26, 2022 |
| Publication Date | March 29, 2022 |
| Submission Date | September 10, 2021 |
| Published in Issue | Year 2022 Volume: 6 Issue: 1 |
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