Identification of S.aureus by specific 16S rRNA and detection of mec A gene from clinical samples in patients of Basrah governorate in Iraq.
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Staphylococcus aureus is a serious human pathogen that causes vast range of contagious conditions both in hospitals and community settings. Methicillin resistant S.aureus (MRSA) are oftenly multidrug resistant in hospital and community that resulted in significant mortality and morbidity. Today, the methicillin resistant S.aureus (MRSA) had become endemic in a hospitals worldwide. So, the need to quick diagnosis and identify of (MRSA) by using PCR technique. A total of 150 clinical specimens include: wound swabs, sputum, throat swabs, nasal swabs, pus and urine, that collected randomly from patients suffering of UTI, wound infection and upper respiratory tract infection, who attended the outpatients and inpatients Clinics of Alsadr Teaching Hospital and Al-Shefa General Hospital in Basrah City south of Iraq country. The identification of the methicillin resistant S.aureus (MRSA) from clinical specimens were performed depending on morphological and biochemical assay based positive cultures of S.aureus from clinical samples. Out of 150 clinical specimens, 61(40.66%) of Staphylococcus aureus were isolated and identified by conventional methods, 20 (62.5%) of these bacterial isolates were from ear pus samples followed by 18 (60.0%) were from sputum, 11 (26.82%) were from urine, 6 (28.57%) were from nasal, 4 (21.05%) were from throat and 2 (28.57%) were from wound samples respectively. Also, 56 (37.33%) of (CONS) were detected from clinical samples and they included Staphylococcus epidermidis which was frequently isolated bacterial species 41 (73.21%) followed by 15 (26.78%) of Staphylococcus saprophyticus. All isolates of S.aureus were be examined for methicillin resistance by disc diffusion assay of oxacillin and cefoxitin antibiotics and by using modified Kirby-Bauer method. Inhibition zones according to the Clinical and Laboratory Standards Institue (CLSI, 2021) which recorded a higher resistance (100%) to both antibiotics. PCR based molecular method was used for accurate identification of specific 16S rRNA gene and mec A gene in S.aureus (MRSA) isolates from clinical samples. The results of PCR amplification of 16srRNA specific gene from S.aureus isolates revealed that 52/61 isolates from a total 61 of S.aureus that detected in clinical samples through conventional methods with percentage of (85.24%) had a clear bands of approximately 228 bp which corresponds to identification of S.aureus strains. While, the PCR amplification of 42/52 isolates with percentage of (80.76%) were generated a clear bands of approximately 310 bp which corresponding to detect mec A gene in methicillin resistant strains of S.aureus isolates.
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The reviewers raise concerns regarding the scientific rigour and experimental design of the work.
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Comments to Author
Dear authors, The document presents valuable findings on the identification of S. aureus and MRSA in clinical samples. However, addressing the below comments regarding grammar, missing information, clarity, and limitations will enhance the quality and impact of the research. Grammar Corrections Sentence Structure: Original: "Methicillin resistant S.aureus (MRSA) are oftenly multidrug resistant in hospital and community that resulted in significant mortality and morbidity." Correction: "Methicillin-resistant S. aureus (MRSA) is often multidrug-resistant in hospitals and communities, resulting in significant mortality and morbidity." Punctuation: Ensure consistent use of commas, especially in lists. For example, "wound swabs, sputum, throat swabs, nasal swabs, pus and urine" should have a comma before …
Comments to Author
Dear authors, The document presents valuable findings on the identification of S. aureus and MRSA in clinical samples. However, addressing the below comments regarding grammar, missing information, clarity, and limitations will enhance the quality and impact of the research. Grammar Corrections Sentence Structure: Original: "Methicillin resistant S.aureus (MRSA) are oftenly multidrug resistant in hospital and community that resulted in significant mortality and morbidity." Correction: "Methicillin-resistant S. aureus (MRSA) is often multidrug-resistant in hospitals and communities, resulting in significant mortality and morbidity." Punctuation: Ensure consistent use of commas, especially in lists. For example, "wound swabs, sputum, throat swabs, nasal swabs, pus and urine" should have a comma before "and" for clarity: "wound swabs, sputum, throat swabs, nasal swabs, pus, and urine." Word Choice: Replace "oftenly" with "often" as "oftenly" is not a standard English word. Subject-Verb Agreement: Original: "All isolates of S.aureus were be examined for methicillin resistance..." Correction: "All isolates of S. aureus were examined for methicillin resistance..." Methodology Details: The document should provide a more detailed description of the PCR protocols, including specific temperatures, times, and concentrations of reagents used. This information is crucial for reproducibility. Sample Size Justification: There should be a justification for the sample size of 150 clinical specimens. Explain how this number was determined to be statistically significant. Demographic Data: Include demographic information about the patients, such as age, gender, and any underlying health conditions, which could impact the prevalence of S. aureus. Statistical Analysis: The document lacks details on the statistical methods used to analyze the data. Specify the statistical tests applied and the rationale for their use. Ethical Considerations: Mention whether ethical approval was obtained for the study and how informed consent was secured from the patients. Experimental Design: The rationale behind the selection of clinical samples (e.g., wound swabs, sputum) should be clarified. Why were these specific samples chosen, and how do they relate to the study's objectives? Results Presentation: The results section could benefit from clearer organization. Consider using tables or figures to summarize the data on the prevalence of S. aureus and MRSA in different sample types. Discussion of PCR Results: The document mentions the amplification of specific gene sequences but lacks a discussion on the significance of these findings. Explain how the presence of these genes correlates with clinical outcomes. Sample Diversity: The study may be limited by the geographic location (Basrah, Iraq) and the specific hospitals involved. This could affect the generalizability of the findings to other regions or populations. Potential Biases: Discuss any potential biases in sample collection, such as selection bias or reporting bias, which could influence the results. Resistance Mechanisms: The study focuses on the mecA gene but does not explore other potential resistance mechanisms. Acknowledging this limitation would provide a more comprehensive understanding of MRSA. Follow-Up Studies: The document should mention the need for follow-up studies to confirm the findings and explore the clinical implications of MRSA prevalence in the studied population. Impact of Antibiotic Use: Discuss how the misuse of antibiotics in the community may contribute to the emergence of MRSA, as this is a critical factor in understanding the resistance patterns observed.
Please rate the manuscript for methodological rigour
Poor
Please rate the quality of the presentation and structure of the manuscript
Poor
To what extent are the conclusions supported by the data?
Partially support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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