ABSTRACT
Staphylococci are the most frequently isolated bacteria from blood. The prevalence of antibiotic resistance has dramatically increased. Real-time PCR offers rapid, accurate, and sensitive method to detect the presence of antimicrobial resistance. This study is mainly aimed to detect uethicillin [oxacillin] and vancomycin resistant staphylococci isolated from blood of patients with significant bacteremia in Assiut University Hospital using both real-time PCR and phenotypic agnostic methods. Sixty Staphylococcal isolates were included. These isolates were collected from positive blood culture bottles [BACTEC 9050 System] of patients with significant bacteremia in Assiut University Hospital. Identification of staphylococcal species was performed by subculture on Mannitol Salt Agar [MSA] and by Microscan system, while antibiotic sensitivity testing was performed by Microscan system, Epsilometer test [E- test], Disc diffusion [DD] method, Oxacillin Resistance Screening Agar Base [ORSAB] and by real-time PCR for mec A and van A genes. Seven S. aureus isolates and fifty three Coagulase-negative staphylococci [CoNS] were detected by both MSA and Microscan system. The most effective antibiotics for staphylococcal isolates were in order. Vancomycin, Linezolid, Synercid, Rifampicin, Chloramphenicol, Gentamycin, Tetracycline, Trimethoprim/Sulphamethoxazole, Clindamycin, Ofloxacin, Ciprofloxacin, Azithromycin and Erythromycin. Concerning methicillin resistance, Real-time PCR which is the gold standard method detected mecA gene in 57 isolates. Accordingly, sensitivity and specificity of E-test were 964% and 100% respectively, DD method showed 87.7% sensitivity and 100% specificity, ORSAB media showed 92.9% sensitivity and 100% specificity while Microscan showed 100.0% sensitivity and 100.0% specificity. Concerning vancomycin resistance, E-test which is the gold standard method detected vancomycin resistance in 6 staphylococcal isolates. Therefore, the DD method showed 66.7% sensitivity and 100.0% specificity while Microscan showed 83.3% sensitivity and 100.0% specificity. Real-time PCR detected van A gene in only one staphylococcal isolate. CoNS organisms are more implicating than S. aureus in bloodstream infections [BSIs]. About 95% of staphylococcal isolates were resistant to methicillin and 10% were resistant to vancomycin. Real-time PCR was more accurate and rapid method for detection of methicillin resistance than phenotypic methods and it could be considered a confirmatory method for detection of vancomycin resistance in staphylococcal isolates suspected to have the van A gene
Subject(s)
Vancomycin/genetics , Polymerase Chain Reaction/methods , PhenotypeABSTRACT
Diabetes is important as a cause of cardiovascular disease [CVD], ranging from asymptomatic ischemia to clinically evident heart failure. Therefore, early identification of sub-clinical CVD in diabetic patients may be particularly important in leading to early initiation of treatment. The aim of the present study was to identify role of BNP [Brain natriuretic Peptide]. Ankle Brachial Index [ABI] and carotid Doppler in detection of sub-clinical CVD in type 2 diabetic patients. BNP was measured in 60 consecutive diabetic patients [patients group] whom were attended internal medicine outpatient clinics or admitted at endocrinology unit of Assiut university hospital. Another 40 patients; were chosen as [control group] their age and sex matched with patients. Echocardiography examinations were performed to all participants. ABI measurements were eonducted on all study participants. Carotid intima Media Thickness [CIMT] and carotid Plaque were evaluated by Carotid Doppler Ultrasonography, along with the determination of anthropometric parameters, HbAlc, lipid profile, assessment of diabetic retinopathy, nephropathy, and neuropathy, in patients with type 2 diabetes mellitus [T2DM]. Our study revealed 11 patients had Left ventricular hypertrophy [LVH], 20 patients had Left Ventricular Diastolic Dysfunction [LVDD], and no systolic dysfunction were detected. BNP were independent determinants of mild to moderate LVDD. Prevalence of a low ABI [<0.9] was 18.3%. Patients with low ABI had significant increased mean ages [P=0.038] duration of DM [P=0.004], concentration of HbAlc [P=0.044], BNP [P=0.013] and microalbuminurea [P-0.007].Patients with low ABI significantly associated with nephropathy [P=0.001], retinopathy [P=0:007], LVH [P=0.010] LVDD [P=0.018] and carotid artery atherosclerosis [P=0.018]. 20 patients [33.3%] were found to have evidence of carotid artery disease of them 5 patients [8.3%] had increased CIMT and 15 patients [25%] had carotid artery plaques, patients with carotid plaque were significantly smoker [P=0.008], male gender [P=0.013], had low HDL [P-0.008] and higher concentration of HbAlc [P=0.001]. Also patients with carotid artery atherosclerosis were significantly associated with nephropathy [P=0.000], neuropathy [P=0.050], Peripheral Arterial Disease [PAD] [P=0.018], LVDD [0.002].Conclusion: Our study showed that BNP discriminated patients at high risk for mild to moderate LVDD. A low ABI were prevalent in our study and associated with age, duration of diabetes, high HbAlc, microalbuminurea and chronic complication of DM, also carotid atherosclerosis high prevalent in our study especially carotid plaques which significantly associated with male gender, smoking, high HbAlc, low HDL, LVDD, PAD, and diabetic microangiopathy