Detection of Biofilm in Surgical Site Infection by Microtiter Plate Method and Its Correlation with icaD, and icaA genes in Staphylococcus Spp

M. I. El-Amir, Asmaa Gaber, Mohamed Yousef, T. Gamal, M. A. El-Feky
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Abstract

Purpose: To determine the bacteriological pattern of bacteria causing surgical site infection (SSI) in Qena University Hospitals and detection of biofilm producing organisms. Methodology: Wound swab from 114 patients with SSI were collected for identification of bacteria and detection of biofilm producing organisms by microtiter plate (MTP) method and tube method (TM). icaA and icaD genes were detected in Staphylococcus spp. by polymerase chain reaction (PCR) from extracted bacterial DNA Principal Findings:Patients mean age was 33.6 ± 12.7 years. 56.1% of the patients were enrolled to emergent surgeries. Diabetes mellitus was the most common risk factor detected in 17.5%. The highest infection rates were noticed after appendectomy, then followed by cholecystectomy (45.6% and 36.8% respectively). Most of bacteria isolated were gram-negative bacteria 67.2%. Escherichia coli was the most frequent bacteria (32.7%), followed by Klebsiella pneumonia (26.2%), Staphylococcus aureus (22.9%), Staphylococcus epidermidis (9.8%), Pseudomonas aeruginosa(5.7%), then Proteus spp (2.5%). Significant biofilm production were detected between Staphylococcus aureus and E. colispp by MTP method (P value Staphylococcus strains, While all Staphylococcus strains were negative for icaA gene. All positive strains for icaD gene were strong biofilm producer by MTP method. Conclusion: MTP was better than TM in detection of biofilm formation. icaD gene were positive in strong biofilm producer Staphylococcus by MTP method while icaA not.
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微滴板法检测手术部位感染生物膜及其与葡萄球菌icaD、icaA基因的关系
目的:了解Qena大学附属医院手术部位感染病原菌的菌型及产膜菌的检测情况。方法:收集114例SSI患者创面拭子,采用微滴板法(MTP)和试管法(TM)进行细菌鉴定和产膜微生物检测。采用聚合酶链反应(PCR)对葡萄球菌进行icaA和icaD基因的检测。主要发现:患者平均年龄33.6±12.7岁。56.1%的患者被纳入紧急手术。糖尿病是最常见的危险因素,占17.5%。术后感染率最高的是阑尾切除术,其次是胆囊切除术,分别为45.6%和36.8%。检出的细菌以革兰氏阴性菌居多,占67.2%。最常见的细菌是大肠杆菌(32.7%),其次是肺炎克雷伯菌(26.2%)、金黄色葡萄球菌(22.9%)、表皮葡萄球菌(9.8%)、铜绿假单胞菌(5.7%),其次是变形杆菌(2.5%)。MTP法检测到金黄色葡萄球菌和大肠杆菌之间有显著的生物膜生成(P值葡萄球菌),而所有葡萄球菌的icaA基因均为阴性。所有icaD基因阳性菌株均为MTP法强生膜菌。结论:MTP检测生物膜形成优于TM。MTP法在强生膜葡萄球菌中检测到icaD基因阳性,而icaA基因不阳性。
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