Background. Neonatal septicemia is a critical medical situation; current conventional laboratory methods still have many limitations and diagnostic obstacles. For this purpose, last decades have witnessed a challenge between the battery of sepsis biomarkers including many leukocyte surface antigens, not only for early diagnostic purposes but also for better follow-up and good management of sepsis patients. Aim. To evaluate the diagnostic, prognostic, and monitoring performance of both neutrophil CD64 (nCD64) and presepsin as sepsis biomarkers compared to each other and to the conventional laboratory sepsis parameters aiming to decide which is the best fitting for routine daily use in neonatal intensive care units (NICUs). Methods. 235 neonates were enrolled from three Egyptian neonatal ICUs, during the period from November 2015 till March 2018; they were classified into two main groups: the control group (n = 102) and the sepsis group (n = 133). Laboratory sepsis evaluation included highly sensitive CRP (hs-CRP), CBC, in addition to nCD64 (flow cytometry technique), and presepsin measurement (CLEIA technique combined with Magtration® technology); the diagnosis was confirmed thereafter by positive blood culture results (BacT/Alert system). Sixty-two of the enrolled sepsis neonates were subjected to follow-up assessment; they were reclassified according to their clinical improvement at the second time assessment into (group 1: sepsis group without improvement) (n = 20) and (group 2: improved sepsis group) (n = 42). Results. Significant increase in nCD64 and presepsin values was found in sepsis groups compared to the controls. At cutoff 41.6%, nCD64% could discriminate the presence of septicemia with sensitivity 94.7%, specificity 93.6 %, and AUC 0.925, while presepsin at cutoff 686 pg/ml achieves sensitivity 82.7%, specificity 95.5%, and AUC 0.887, respectively. Significant increase in nCD64 ( P < 0.001 ) and hs-CRP ( P = 0.018 ) values was observed in severe sepsis/septic shock patients compared to nonsevere sepsis patients. Delta change percentage (dC%) between initial and follow-up evaluations for both improved and nonimproved sepsis patients was dC Z value −5.904 for nCD64% followed by dC Z value −4.494 for presepsin. Conclusion. nCD64 and presepsin are valuable early diagnostic and monitoring sepsis biomarkers; the highest sensitivity could be achieved by a univariant sepsis marker in this study was recorded by the nCD64% biomarker, while the highest specificity was documented by presepsin. Combined measurement of both achieves the highest diagnostic performance in sepsis neonates. Either of CD64 or presepsin combined with hs-CRP associated with better performance than any of them alone. nCD64 carries an additional promising role in reflecting sepsis prognosis.

AIMS:To evaluate three in vitro phenotypic methods; tissue culture plate, tube method, and Congo red agar for detection of biofilm formation in staphylococci and assess the relation of biofilm formation with methicillin resistance and anti-microbial resistance.METHODS:The study included 150 staphylococcal isolates. Biofilm detection in staphylococci was performed using tissue culture plate, tube method, and Congo red agar.RESULTS:Tissue culture plate, tube method, and Congo red agar detected 74%, 42.7%, and 1.3% biofilm producing staphylococci respectively. S. aureus isolates were more common biofilm producers (53.2%) than CONS (46.8%). Biofilm production in CONS species was highest in S. hemolyticus (57.7%). Tube method was 51.4% sensitive, 82.1% specific. As for Congo red agar, sensitivity was very low (0.9%), but specificity was 97.4%. Biofilm producers were mostly; isolated from blood specimens and detected in methicillin-resistant strains 96/111 (86.5%). They were resistant to most antibiotics except vancomycin and linezolid.CONCLUSIONS:Tissue culture plate is a more quantitative and reliable method for detection of biofilm producing staphylococci compared to tube method and Congo red agar. Hence, it can still be used as a screening method for biofilm detection. Vancomycin and Linezolid are the most sensitive antibiotics among biofilm producing staphylococci.

AIM:To evaluate the diagnostic performance of MDI and temocillin disk (30 μg) for detection of carbapenem-resistant Enterobacteriaceae in comparison to real-time PCR.MATERIAL AND METHODS:Fifty specimens submitted to the Microbiology Laboratory of Ain Shams University Hospitals and showed resistance to carbapenem drugs through routine culture and susceptibility testing, were assessed by both temocillin disk (30 μg) and MDI set to detect carbapenem-resistant Enterobacteriaceae. Results were compared to real-time PCR for detection of carbapenemase genes blaKPC, blaNDM, blaOXA–48-like, blaVIM, and blaIMP.RESULTS:Our work revealed that most of the CPE isolates were Klebsiella species (62%) followed by E. coli (24%), Serratia (10%) and Citrobacter (4%). Phenotypic detection of carbapenem-resistant classes revealed OXA - 48 in 96% of isolates, followed by MBLs (82%), and KPC (34%). All isolates were negative for AmpC. Detection of the genes by real-time PCR showed that the predominance was for the blaOXA-48 gene (96%) then blaVIM (94%) followed by blaNDM (54%), blaKPC (46%) and finally blaIMP (40%). Evaluation of the MDI set against PCR showed sensitivity (82.1%) and specificity (70%). The temocillin disk had 97.9% sensitivity and 50% specificity. The evaluation of Temocillin disk and MDI in combination for detection of carbapenem-resistant Enterobacteriaceae showed 99.7% sensitivity and 35% specificity.CONCLUSION:Adding Temocillin disk to Mastdisks ID inhibitor combination set provides a simple, easy, rapid and highly sensitive test that can be used for screening and classification of carbapenem-resistant Enterobacteriaceae. However, it still needs confirmation by molecular techniques.

Background: Patients with end stage renal disease (ESRD) have elevated levels of inflammatory mediators including Creactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). Neutrophil to lymphocyte ratio was introduced as a novel inexpensive indicator that reflects the severity and extension of systemic inflammation and atherosclerosis, and predicts adverse clinical outcomes in cardiac and non-cardiac disorders including ESRD.The aim of this study is to evaluate the frequency of subclinical inflammation using neutrophil to lymphocyte ratio (NLR) and its relation to other biochemical parameters in ESRD patients on maintenance haemodialysis (HD) in comparison to high-sensitivity C-reactive protein (hsCRP). Methods:A cross sectional observational study including 100 clinically stable ESRD patients on regular haemodialysis (mean age 48.25 ± 13.67, 57% were males), NLR was calculated from complete blood count and hsCRP was recorded for all patients where 8.2 mg/L indicated inflammation.Results: NLR was positively correlated to hsCRP, using ROC curve (AUC=0.647) and the best cut off point to detect subclinical inflammation in HD patients was calculated with NLR greater than or equal 1.54, with sensitivity 68.25% and specificity 54%. By using univariate and multiple variate analysis in our work to test for independent predictors of hsCRP levels as an indicator for inflammation, it was found that NLR can be used as an independent predictor of hsCRP as an inflammatory marker, with statistically significant correlation (p=0.015). Spearman coefficient (rs=0.220) shows statistically significant (p=0.028) positive correlation between NLR and hsCRP. Conclusion:NLR is an easy, simple and non-expensive method that can be used as a marker of inflammation in HD patients when compared to hsCRP.

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