Electronic nose versus VITEK 2 system for the rapid diagnosis of bloodstream infections.

Infectious diseases that spread through the bloodstream, known as bloodstream infections (BSIs), are a major global health problem. Positive outcomes for patients with sepsis are typically the result of prompt treatment started after an early diagnosis of BSIs. In this study, we evaluated the capabilities of a portable electronic nose (E-Nose) to detect BSIs with two commonly isolated Gram-negative bacterial species, E. coli and K. pneumonia. One hundred and five blood samples were randomly collected for blood culture examinations using BACTEC and VITEK 2 system, and headspace analysis by an E-Nose from June to December 2021. Classification accuracy of E. coli, K. pneumonia, and negative controls was measured using principal component analysis, area under the receiver operating characteristic curve, sensitivity, and specificity analysis. After incubation for 24 h, cluster plots generated using principal component analysis demonstrated that E-Nose could accurately diagnose the presence of E. coli and K. pneumonia in BACTEC blood culture bottles with a sensitivity and specificity of 100% in just 120 s. The E-Nose method has been shown to be an immediate, precise, and cost-effective alternative to automated blood culture BACTEC and VITEK 2 systems for the fast detection of the causative bacterial pathogens of BSIs in clinical practice. Thus, patients with such Gram-negative bacteremia can have guided empirical antimicrobial therapy on the same day of BSIs diagnosis, which can be lifesaving.

Possible roles of methylenetetrahydrofolate reductase polymorphism and folate status in patients with early hepatitis C virus genotype 4.

BACKGROUND AND STUDY AIMS: Hepatitis C virus (HCV) infection is a major cause of chronic liver disease worldwide. In Egypt, 92.5% of HCV infection cases reportedly involve infection with HCV genotype 4. HCV infection may induce liver steatosis directly and indirectly. Host genetic polymorphisms may also contribute to the pathogenesis of steatosis. Folate deficiency indirectly cuase liver damage. Folate status is mostly affected by MTHFR C677T polymorphism. The pathophysiology of thrombocytopenia (TCP) in HCV infection remains unclear. Thus, the present study investigated the roles and consequences of MTHFR C677T SNP and folate status in patients with early HCV genotype 4 infection and their relation with steatosis and thrombocytopenia. PATIENTS AND METHODS: Fifty patients with the HCV genotype 4 and 50 healthy controls were enrolled in the study. All the participants underwent laboratory, demographic, and anthropomorphic examinations. Serum folate level was determined, and genomic analysis of MTHFR C677T SNP was performed. RESULTS: No significant difference in allelic frequency of MTHFR C677T was observed between patients and controls. However, significantly lower serum folate level, hemoglobin level, and platelet count were found in patients than controls (p = 0.014, p = 0.005, and p = 0.001, respectively). The cholesterol, triglyceride, and high-density lipoprotein levels were also significantly lower in patients than controls (p < 0.001, p = 0.001, and p < 0.001, respectively), whereas the low-density lipoprotein level was significantly higher in patients (p < 0.001). Patients harboring the MTHFR CT genotype had a significantly lower serum folate level (p = 0.033) than the controls. Among the patients with HCV infection, those with the TT genotype had the highest body mass index (p = 0.003) and levels of cholesterol, triglyceride, and high-density lipoprotein (p = 0.007, p = 0.025, and p = 0.040, respectively). CONCLUSION: MTHFR C677T SNP may contribute to the development of complications associated with early HCV genotype 4 infection, such as dyslipidemia and decreased folate levels.

Blood group antigen-Binding Adhesion2 (BabA2) gene in gastric tissue biopsies as a diagnostic biomarker for Helicobacter pylori infection.

BACKGROUND: The Lewis (b) blood group antigen-Binding Adhesion2 (BabA2) has been reported to mediate the attachment of H. pylori to human. AIM: assessment the diagnostic potential of detection of (BabA2) gene compared with immunostaining of Lewis (b) by specific mouse monoclonal antibodies in gastric biopsies from Egyptian Patients as a diagnostic maker for Helicobacter pylori infection. MATERIALS AND METHODS: Fifty untreated patients suffering from dyspeptic complaints were enrolled in this study and underwent for upper gastro-duodenal endoscopy. Biopsies were taken for histological examination by (H&E) and immunohistochemical analysis for Lewis b by specific mouse monoclonal antibodies, and scoring of Lewis b expression in gastric tissue biopsy as well as molecular detection of BabA2 gene of H. pylori by PCR. Biochemical analysis was performed to detect the presence of H. pylori urease activity using Rapid Urease Test (RUT). RESULTS: Out of 50 gastric biopsies, 41 biopsies were positive for histological, Immunostaining for Lewis b expression and urease activity test (RUT) for H pylori. RUT showed a sensitivity of 87.8%, specificity 88.9%, positive predictive value (PPV) 97.2%, and negative predictive value (NPV) 61.5%. BabA2 gene results revealed that, out of 41 positive biopsied cases, 39 (95.1%) were positive by the PCR test for BabA2 gene. And all 9 negative biopsies (100%) for H pylori negative for BabA2gene so the sensitivity and specificity of BabA2 gene detection in gastric biopsies by PCR were 95.1% and 100%; respectively. CONCLUSION: BabA2 gene detection in gastric tissue biopsies could be suggested as a diagnostic biomarker to be included among the other biomarkers routinely performed for clinical diagnosis of H. pylori infection.

Telomerase activity, cytokeratin 20 and cytokeratin 19 in urine cells of bladder cancer patients.

AIM OF THE STUDY: This work aims to search for markers suitable for the screening of bladder cancer, which should be specific, sensitive, reproducible, non-invasive and at acceptable cost. PATIENTS AND METHODS: The study included 50 patients diagnosed as bladder cancer (35 TCC, 15 SCC) of different stages and grades, 30 patients with various urothelial diseases, besides 20 apparently healthy subjects of matched age and sex to the malignant group. A random midstream urine sample was collected in a sterile container for the determination of telomerase by RT-PCR, keratin 19 by ELSA CYFRA 21-1 IRMA kit, keratin 20 by RT-PCR and immunohistochemical staining, and urine cytology. RESULTS: For all parameters (telomerase, K19, K20 and cytology) the malignant group was significantly different from both the benign and the control groups. None of the four studied parameters was correlated to the stage of the disease, and when it comes to grade, only K19 showed a significant positive correlation with grade both in TCC and SCC. When ROC curves for all parameters were compared, K19 had the largest area under the curve, and then comes K20. CONCLUSION: K 19 may be used as a biological marker for the diagnosis of bladder cancer. K19 could not be used for differential diagnosis of different types of bladder cancer, meanwhile it could be a marker for differentiation that decreases in less differentiated tumors. As a tumor marker, K20 reflects inability to differentiate tumor type or grade in TCC, while in SCC of the bladder it is correlated with the grade. As a method, RT-PCR is superior to immunostaining for the detection of bladder cancer, meanwhile K20 immunohistochemistry (IHC) results were much better than urine cytology as a bladder cancer screening test. Haematuria and inflammation reduced the specificity of telomerase assay, which reduced its validity as a tumor marker of bladder cancer. K19 and K20 are the best candidates as screening tests for the diagnosis of bladder cancer, representing the highest sensitivity and specificity, beside the radiological and histopathology. Meanwhile, telomerase, although it was a sensitive enough marker, it reflected a high false positive rate.