Evaluation of a multiplex polymerase chain reaction for the diagnosis of infectious diarrhea in intensive care unit patients in Upper Egypt.

The rapid diagnosis of infectious diarrhea is lifesaving for intensive care unit (ICU) patients. This study evaluated a commercially available multiplex polymerase chain reaction (PCR) (BioFire FilmArray) for the diagnosis of parasitic and bacterial infections in ICU patients with secretory diarrhea in comparison to other traditional methods. This cross-sectional study included 50 subjects with infectious diarrhea. Their stool samples were subjected to macroscopic and microscopic examinations, concentration techniques, permanent staining techniques, stool culture, identification of bacterial infection by the Vitek 2 Compact 15 System, and molecular diagnosis of bacterial or parasitic infections by BioFire FilmArray multiplex PCR. Parasitological examination showed that the sensitivity and specificity of BioFire FilmArray multiplex PCR in the diagnosis of Cryptosporidium oocysts were 83.33% and 100.0%, respectively compared with 100% and 92.5% in diagnosis of G. lamblia cysts. Bacteriological examination showed that the sensitivity and specificity of BioFire FilmArray multiplex PCR in the diagnosis of E. coli and salmonella were 100% and 100.0%, respectively. The BioFire FilmArray multiplex PCR gastrointestinal (GI) panel assay was more sensitive and specific in the diagnosis of bacterial infections than parasitic infections. The BioFire FilmArray multiplex PCR GI panel assay was less sensitive in the detection of Cryptosporidium oocysts than traditional methods. In conclusion, the BioFire FilmArray multiplex PCR may be useful for rapid diagnosis of ICU patients with infectious diarrhea.

Inetrleukin-6 and C-reactive protein as predictors of mortality among critically ill COVID-19 patients in Assiut university hospitals (ICUs).

COVID-19 represents a serious global threat due to scarcity of definitive cure and its infectious nature. The death rate of COVID-19 patients admitted to hospitals was quite high, and cytokine storms could be the mechanism of severity. Interleukin-6 (IL6) and C-reactive protein (CRP) may predict severity and mortality. We attempted to determine the role of IL6 and CRP as predictors of death in intensive care unit (ICU) patients. This Cross-sectional hospital study included 100 patients admitted to ICUs at Assiut University Hospitals from October 2020 to October 2021. Data including age, sex and comorbidities were recorded, laboratory investigations included CRP, ferritin, and IL6. Data were collected and analyzed. Morality predictors in ICU patients with COVID-19 infection were older age (>60 years), presence of diabetes mellitus, chest diseases, CRP >49, IL-6 >70 pg/ml. In conclusion, early ranking and identification of people, who are at risk of death among ICU patients, by monitoring of CRP, IL6, early treatment of cytokine storm, and good management of pre-existing comorbidities would be a very useful approach to reduce the mortality among ICU patients.

Genetic detection of AmpC beta-lactamase among gram negative isolates "A Single Center Experience''.

The misuse of ß-Lactam antibiotics results in major problem, microbial resistance against these antibiotics by expression of ß-lactamases, facing its use. AmpCs are one of the ß-lactamases which confer resistance to penicillins, cephalosporins, cephamycins, and aztreonam, and are not affected by classic ß-lactamase inhibitors. Plasmid-mediated AmpC ß-lactamases pose a major challenge to infection control because the AmpC gene can be expressed in larger quantities and has a high transmissibility to other bacterial species. This study aimed to detect plasmid mediated AmpC ß-lactamases in gram negative isolates in Assiut university hospital. It was performed on 120 cefoxtin resistant isolates obtained from 300-gram negative isolates using the disc diffusion method as a screening test for AmpC production. Since the presence of pAmpC is often associated with the presence of ESBLs, phenotypic detection of ESBL was done using combined disc method and vitek2 compact 15. Phenotypic detection of AmpC was done by disc approximation method and inhibitor-based method using phenyl boronic acid (PBA). Genotypic detection of 5 plasmid mediated AmpC genes families (MOX, CIT, DHA, EBC, and FOX) was done by multiplex PCR. Our result showed that Klebsiella pneumoniae (62.5%) and Escherichia coli (25.8%) were the most frequent isolates. Only 15.8 %, 12.5%, 17.5% resistant isolates to cefoxitin were positive by using disc approximation test, inhibitor-based method using PBA (150 g/mL), and PBA (600 g/mL), respectively. Out of the 120 Cefoxitin-resistant isolates, 22 isolates (18.3 %) were positive by multiplex PCR. CIT and MOX were solely detected in 45.5% and 4.5%, respectively. CIT and FOX together were detected in 45.5%, CIT and DHA together in 4.5%. No isolate was positive for EBC gene. Finally, boronic acid test using 600 µg/mL PBA with 30µg ceftazidime, as phenotypic method for detecting AmpC ß- lactamases, was ranked very good for marking negative tests.

Epidemiology of Carbapenem-resistant Enterobacteriaceae in Egyptian intensive care units using National Healthcare-associated Infections Surveillance Data, 2011-2017.

Objective: To describe the epidemiology of carbapenem-resistant Enterobacteriaceae (CRE) healthcare-associated infections (HAI) in Egyptian hospitals reporting to the national HAI surveillance system. Methods: Design: Descriptive analysis of CRE HAIs and retrospective observational cohort study using national HAI surveillance data. Setting: Egyptian hospitals participating in the HAI surveillance system. The patient population included patients admitted to the intensive care unit (ICU) in participating hospitals. Enterobacteriaceae HAI cases were Klebsiella, Escherichia coli, and Enterobacter isolates from blood, urine, wound or respiratory specimen collected on or after day 3 of ICU admission. CRE HAI cases were those resistant to at least one carbapenem. For CRE HAI cases reported during 2011-2017, a hospital-level and patient-level analysis were conducted using only the first CRE isolate by pathogen and specimen type for each patient. For facility, microbiology, and clinical characteristics, frequencies and means were calculated among CRE HAI cases and compared with carbapenem-susceptible Enterobacteriaceae HAI cases through univariate and multivariate logistic regression using STATA 13. Results: There were 1598 Enterobacteriaceae HAI cases, of which 871 (54.1%) were carbapenem resistant. The multivariate regression analysis demonstrated that carbapenem resistance was associated with specimen type, pathogen, location prior to admission, and length of ICU stay. Between 2011 and 2017, there was an increase in the proportion of Enterobacteriaceae HAI cases due to CRE (p-value = 0.003) and the incidence of CRE HAIs (p-value = 0.09). Conclusions: This analysis demonstrated a high and increasing burden of CRE in Egyptian hospitals, highlighting the importance of enhancing infection prevention and control (IPC) programs and antimicrobial stewardship activities and guiding the implementation of targeted IPC measures to contain CRE in Egyptian ICU's .

Glypican-3 and Melanoma Antigen Genes 1 and 3 as Tumor Markers for Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is the commonest liver cancer; its incidence and prevalence are continuously increased. Glypican3 (GPC3), melanoma antigen-1, 3 genes (MAGE1 and 3) are tumor markers used in HCC. We evaluated their role in HCC detection and assessed their relation to tumor parameters. Three groups, HCC group, liver cirrhosis group and a control group were studied. AFP, GPC3, and MAGE1 and 3 mRNA were determined in all study subjects. Tissue GPC3 was examined in patients with HCC only. Serum AFP and GPC3 were elevated in HCC group compared to other groups (P < 0.000 and P < 0.001, respectively). AFP at cutoff 44.4ng/ml and GPC3 at cutoff 5.6µg/L resulted in 81% and 90.1% sensitivity, 73.3% and 92.6% specificity, respectively. The combined measurement of both increased the sensitivity and the specificity to 100% and 93.3%, respectively. GPC 3 was detected in tissues of 81.0% of the cases. MAGE-1 and MAGE-3 genes expression were detected in 61.9% and 52.4%, respectively in HCC cases but not in other groups. GPC3, MAGE1and 3 were increased with advanced tumor stage, size, and nodule numbers. We concluded that GPC3 is a promising diagnostic marker for HCC, and MAGE 1 and 3 could be helpful in early detection of extrahepatic metastasis of HCC.

Hepatitis C infection, Cognition, and inflammation in an Egyptian sample.

Chronic hepatitis C (HCV) infection is associated with cognitive impairments which might be mediated through a secondary inflammatory cascade. Egypt has an unusually high prevalence of HCV monoinfections and is an ideal site for the study of the isolated effects of HCV infection. Therefore, in a hospital-based cross-sectional study based in Egypt, this study compared cognitive functioning and serum markers of inflammation in 11 HCV positive cases and 14 HCV negative controls. The Wisconsin Card Sorting Test was used to assess cognitive flexibility and the Brief Visuospatial Memory Test-Revised was used to assess learning and memory. Circulating levels of soluble tumor necrosis factor receptor II (sTNFR-II), monocyte chemotactic protein-1 (MCP-1/CCL2), and soluble CD14 (sCD14) were determined as indices of inflammation. HCV positive cases had higher levels of sTNFR-II (t = -3.5, P = 0.002). HCV positive cases also had significantly worse cognitive flexibility with higher number of total errors (t = -2.18, P = 0.04), and preservative responses (t = -2.12, P = 0.05), and lower number of conceptual level responses (t = 1.32, P = 0.04) on the Wisconsin Card Sorting test. In conclusion, results from this pilot study indicate that HCV+ patients have worse cognitive performance and somewhat greater inflammatory activity as compared to controls. The increased inflammation may be associated with the cognitive impairments observed in these HCV+ patients.

Clinico-laboratory study on children with auto-immune hepatitis in Upper Egypt.

BACKGROUND AND STUDY AIMS: Auto-immune hepatitis (AIH) in children is a rare chronic progressive liver disorder. It is characterised serologically by high aminotransferase levels, elevated immunoglobulin G (IgG) and the presence of autoantibodies. AIH is divided into two types according to the autoantibody profile. This study aims to assess frequency, clinical manifestations, biochemical features and outcome of AIH in children attending Assuit University Hospitals in Upper Egypt with acute icteric hepatitis and seronegative viral markers (anti-hepatitis A virus (HAV) IgM, HbsAg, anti-hepatitis C virus (anti-HCV) Ab). PATIENTS AND METHODS: The study includes 34 children with AIH, diagnosed on the basis of the International Scoring Criteria of Auto-immune Hepatitis, recruited from Assuit University Hospitals, during the period from January 2005 to December 2009. All patients received prednisolone 2mgkg(-1)day(-1). Follow-up was done for 1year. RESULTS: Among 34 children diagnosed as AIH, 24 were females (70.5%) and 10 were males (29.5%). Jaundice represented the most consistent finding in all patients. According to the autoantibody profile, 25 children were classified as type 1 and nine children were classified as type 2. Corticosteroid therapy was started. Complete remission was observed in 67.6% of patients and partial remission in 17.6%. There was no significant statistical difference in clinical and biochemical features of AIH in patients regarding the response to treatment. Mild side effects of steroid therapy were encountered in 48.2% of patients. After complete withdrawal of corticosteroids, six patients (20.7%) developed relapse. CONCLUSION: AIH type 1 was the main form of AIH in children referred to Assiut University Hospitals. Girls were more affected than boys. AIH type 1 exhibited a more active, ongoing immunologic process. Steroid alone can be used successfully in most cases. Children with AIH type 2 had a higher frequency of relapse after corticosteroid withdrawal. Further studies on a larger number of cases and long-term follow-up are recommended.

Clinicopathologic features and genotyping of patients with chronic HBV infection in the Upper Egypt.

The aim of this study was to determine the clinicopathologic features and Hepatitis B virus genotypes in HBV-infected patients in the Upper Egypt. Eighty-three HBsAg-positive patients (28 carriers, 14 with chronic hepatitis, 32 with liver cirrhosis and 9 with hepatocellular carcinoma) were enrolled. Blood was collected and serum samples obtained were screened for Hepatitis markers genotyping was conducted for 6 HBV genotypes (A through F) using a method for genotyping HBV by primer specific polymerase chain reaction. Genotype D was the only genotype detected in different clinical forms of chronic HBV infection (carriers, chronic hepatitis, cirrhosis and hepatocellular carcinoma) and, in all patients who had elevated or normal alanine aminotransferase levels and in all ages. HBeAg was absent in 78 patients suggesting the presence of pre-core or core mutations. Positive correlation was found among serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), histological activity index and grade of hepatitis. This study provides the first indication about the clinicopathologic features of HBV-infected patients in the Upper Egypt. It also reports the predominance of genotype D in this region.

Comparison of different phenotypic and molecular methods for detection of methicillin-resistant Staphylococcus aureus in intensive care patients.

Detection of methicillin-resistant Staphylococcus aureus (MRSA) has been problematic ever since its discovery. This work was performed to evaluate the effectiveness of different phenotypic methods for MRSA detection in intensive care patients. Three hundred and eighty-nine specimens from 100 patients were inoculated onto mannitol salt agar. All S. aureus isolates were examined by four selective culture media [ORSAB, MSO, MSA-Cefox, OAS], two disc diffusion methods [CDD and ODD], and MicroScan panel for MRSA. Polymerase chain reaction (PCR) for mecA gene was performed as the gold standard. S. aureus isolates were revealed from 56 patients, 41 of them were found to be MRSA by PCR. CDD yielded the best sensitivity (97.6%), followed by ODD and MSA-Cefox (92.7%). CDD, MSA-Cefox and OAS showed the best specificity (100%). In conclusion, MSA-Cefox and CDD showed improved sensitivity and excellent specificity compared to other methods. It is advisable to use the two methods for MRSA detection if PCR is not available.

Is Helicobacter pylori infection associated with alopecia areata?

BACKGROUND: Alopecia areata (AA) is an immune-mediated form of hair loss that occurs in all ethnic groups, ages, and both sexes. Helicobacter pylori has been associated with certain extra-digestive dermatological conditions, including chronic urticaria, rosacea, Schönlein-Henoch purpura, Sweet syndrome, systemic sclerosis, and atopic dermatitis. OBJECTIVE: The causal relation between alopecia areata and H. pylori is discussed. We have screened for the presence of H. pylori in patients with AA in order to determine any potential role in its pathophysiology. PATIENTS AND METHODS: We have prospectively studied 31 patients with AA and 24 healthy volunteers of similar gender for the presence of H. pylori surface antigen (HpSag) in stool. RESULTS: Optical density values for H. pylori infection were positive in 18 of all 31 patients evaluated (58.1%), while in 13 patients, values did not support H. pylori infection (41.9%). While in the control group, 10 of 24 (41.7%) had positive results. Within the group of AA, there was no significant difference between HpSag-positive and HpSag-negative patients. CONCLUSIONS: Based on these results, the relation between H. pylori and AA is not supported. We advise that H. pylori detection should not be included in the laboratory workup of AA.

Diagnostic performance and predictive value of rheumatoid factor, anti-cyclic-citrullinated peptide antibodies and HLA-DRB1 locus genes in rheumatoid arthritis.

BACKGROUND: We evaluated the significance of the genes, defined as DRB1*04 or DRB1*01, in rheumatoid arthritis (RA) patients. We focused on the role of genetic and serologic markers to predict disease activity and destructive process of joints. METHODS: Sixty patients with RA were examined. Radiographic changes were evaluated by (Larsen score) and disease activity was measured by disease activity score 28 (DAS28). The markers analyzed were: erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), anti-cyclic citrullinated peptides (anti-CCP2) and HLA-DRB1 alleles typed by PCR. RESULTS: In this study, anti-CCP antibodies, CRP, RF and AKA were detected in 83.3%, 56.7%, 71.7% and 52% of patients respectively. HLA-DRB1*01 was found in 45% of patients and 35% of them had one or two HLA-DRB1*04 alleles. According to DRB1*04 subtypes, (DRB1* 0405) was present in of 80% them. For prediction of grade of activity, the independent predictors were anti-CCP (OR 19.6), and DRB1*04 positive allele (OR 5.1). The combination of DRB1*04 + anti-CCP antibodies gave increase in the specificity and positive predictive value to 92% and 90 respectively. As regards to the prediction of radiological joint damage, the independent predictors were HLA-DRB1*04, HLA-DRB1*01, RF, and CRP > 18 (OR 5.5, 4.5, 2.5, 2.0 respectively). CONCLUSION: Our findings suggest that anti-CCP2 is superior to RF for the detection of RA and provided predictive information on joint destruction and disease activity. The presence of RA associated antibodies (ACCP or RF) and/or the SE genes are indicative for a poorer radiological outcome and higher grade of activity.

Serum IL-2 and platelet-associated immunoglobulins are good prognostic markers in immune thrombocytopenic purpura.

Immune thrombocytopenic purpura (ITP) is an acquired disease in which autoantibodies to platelets cause their sequestration and destruction by mononuclear macrophages, principally in the spleen. While most children with the disease experience a relatively short and benign clinical course, ITP in adults often lasts more than 6 months (chronic ITP) and is resistant to conventional treatment (corticosteroids, intravenous immunoglobulin, or splenectomy). This work was done to study the immunological difference between acute and chronic ITP, the effect of treatment on the studied immunological parameters, and to evaluate the role of prednisone therapy in chronic ITP. The study included 49 patients, twenty-three children with acute ITP, and twenty-six with chronic ITP. After taking the history, clinical examination was performed for all patients and control subjects. Laboratory investigations included complete blood count, bone marrow aspirate examination (patients), direct and indirect Coombs' test, antinuclear antibodies, lymphocyte phenotyping, cytokine (IL-2, IFN-gamma, and IL-6) measurement, and platelet antibodies by immunofluorescence. Results showed that acute ITP is more prevalent in preschool children and its relapse is lower when steroids are used for treatment. Platelet counts were significantly elevated in both acute and chronic ITP, especially with good response to steroids. Also, CD4 and CD4/CD8 were significantly reduced in chronic ITP with good response to therapy. Both IL-2 and IFN-gamma were significantly increased in chronic ITP when compared to acute ITP or control. Platelet associated IgM was detected more in acute than in chronic ITP, while IgG was equally detectable in both cases. This work shows that IL-2 is a good prognostic factor in chronic ITP and steroids are important for its treatment. It also shows that platelet associated IgG is a good monitoring parameter for response to treatment.