Extended-Spectrum ß-Lactamase-Producing Escherichia coli and Virulence Genes in Pediatric Patients with Health-Care Urinary Tract Infections.

INTRODUCTION: Healthcare-associated urinary tract infection (UTI) represents a significant health problem, especially in infants and young children. The most common pathogen associated with this infection is Escherichia coli (E. coli). OBJECTIVE: The present study aimed to detect the frequency of virulence genes among clinical isolates of E. coli isolated from healthcare-associated urinary tract infections in children and the correlation between these virulence genes and the presence of the blaCTX gene. METHODS: The study included one hundred clinical isolates of E. coli isolated from healthcareassociated urinary tract infections in children in intensive care units. The isolates were subjected to antibiotics sensitivity by disc diffusion method and detection of extended-spectrum beta-lactamase by double disc diffusion method. In addition, multiplex polymerase chain reaction (PCR) was used to detect some virulence genes, and PCR was used to detect the blaCTX-M gene. RESULTS: E. coli producing ESBL by double discs method was identified in 74 isolates. blaCTX-M gene detection by PCR was identified among 38 isolates representing 51.4% of ESBL-producing E. coli. There was a significant association between ESBL and blaCTX-M Gene, P = 0.0001. The frequency of the studied virulence genes by multiplex PCR in the isolated E. coli was 66% for the Fim gene, 75% for the Aer gene, 68% for the FliC gene, 53% for each of IucD gene and Usp gene, 40% for pap gene, 35% for each of AFA and ironN genes and 17% for sfa gene. None of the isolated E. coli had the Cdt gene. There was a significant association between the presence of the FimH gene (P = 0.0001), Pap gene (P = 0.05), sfa (P = 0.026), Afa gene (P = 0.018), and aer gene (P = 0.035) and the presence of the blaCTX-M gene in the isolated E. coli. CONCLUSION: The present study highlights the presence of virulence genes and blaCTX-M gene in uropathogenic E. coli isolated from pediatric patients with healthcare-associated urinary tract infections. There was an association between the blaCTX-M gene and virulence genes FimH, pap, sfa, Afa, and aer. Various distributions of the studied genes with a high frequency of fimbria are flic genes. Moreover, the ESBL had high frequency in E. coli with the presence of blaCTX-M in about one-third of the isolates.

Multiplex PCR: Aid to more-timely and directed therapeutic intervention for patients with infectious gastroenteritis.

BACKGROUND: Multiplex PCR is a sensitive and rapid method compared with conventional methods. Therefore, we use multiplex PCR for the rapid detection of the four major intestinal pathogens causing gastroenteritis (Shigella spp., Campylobacter spp., Aeromonas spp. and Enterohemorrhagic Escherichia coli [EHEC]) in stool specimens. MATERIALS AND METHODS: A prospective randomized study using 200 stool samples obtained from patients presented with acute gastroenteritis during the study period (between February 2019 and December 2021). Bacteria in stool samples were identified using conventional culture methods and multiplex PCR for stool samples. RESULTS: The identified organisms using conventional cultures; were Shigella (27%), Aeromonas species (10%) and EHEC (O157) (8%). Using multiplex PCR. Shigella spp. was the most commonly identified pathogen (detected in 40.5% of positive samples), followed by Aeromonas spp. (30%), EHEC (20%) and Campylobacter species was only detected in (1%) of positive samples. The diagnostic evaluation of multiplex PCR in relation to conventional method in diagnosis of Shigella, EHEC and Aeromonas showed, sensitivity of 100% (for each), specificity of 88.5%, 92.4%, 77.8% respectively. However, the diagnostic evaluation of multiplex PCR in relation to conventional method in diagnosis of Campylobacter showed specificity of 99% and NPV of 100%. CONCLUSIONS: Multiplex PCR is an accurate and rapid method for detection of common intestinal pathogens causing severe gastroenteritis. a rapid method that could be used in outbreaks for diagnosis of the common enteric pathogens causing fatal gastroenteritis.

Viral pathogens of acute gastroenteritis in Egyptian children: role of the parechovirus.

BACKGROUND AND AIM: Human parechovirus (HPeV) has emerged as a pathogen associated with acute gastroenteritis (AGE). AIM: To detect the presence of HPeV in the stool samples from Egyptian children with AGE seeking care and the possibility of its co-infection with other enteric viruses. METHODOLOGY: One hundred stool samples were collected from children attending Mansoura University Children's Hospital with AGE. HPeV and astrovirus were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). At the same time, detection of rotavirus antigen and norovirus was achieved by enzyme-linked immunosorbent assay and rapid immunochromatographic method, respectively. RESULTS: The most frequently detected virus was rotavirus (39%), followed by norovirus (27%), HPeV (19%), and astrovirus (12%). Interestingly, the single infection with HPeV was 5%. Among the 19 HPeV positive samples, the co-infection of HPeV with other enteric viruses was detected in 9(43.9%) for rotavirus, 7(36.8%) for norovirus, 2(10.5%) for astrovirus, in 3(15.8%) for rotavirus and norovirus and 1(5.3%) for norovirus and astrovirus. Regarding the clinical presentation, there was no significant difference between children infected with HPeV alone and those infected with viruses other than HPeV alone; fever (p = 0.3), vomiting (p = 0.12), abdominal pain (p = 0.12), and grades of severity (P = 0.82). HPeV alone infected children were of mild severity (60%), and their main presenting symptom was fever (60%). CONCLUSIONS: Detection of HPeV as a single viral pathogen in the stool of some children with AGE showed that this virus could be a causative agent of AGE in Egyptian children. Therefore, HPeV could be included as one of the viruses screened for AGE diagnosis in children in Egypt.

Infection with adenovirus, rotavirus, and coinfection among hospitalized children with gastroenteritis in an Egyptian university hospital.

Acute gastroenteritis is the main cause of mortality and morbidity in children worldwide. Studies stated that rotavirus and human adenovirus (HAdV) are common causes of nonbacterial gastroenteritis in children aged 0-5 years. The aim of this study was to determine the prevalence and the distribution of rotavirus, HAdV, and coinfections among hospitalized children with gastroenteritis below 7 years old and determine the prevalence of enteric HAdV among all HAdV gastroenteritis. The study was conducted on 150 children below 7 years old. Antigen detection for rotavirus and HAdV by ELISA and determination of enteric HAdV (serotype 40 and 41) by nested PCR and restriction endonucleases study were performed. Detection of rotavirus and HAdV antigens in 150 stool specimens from patients with gastroenteritis were 58% (87), 6.7% (10), and 8% (12) positive for rotavirus, HAdV, and coinfection, respectively. Out of 22 HAdV antigen-positive cases, 15 cases were positive by PCR for enteric HAdV, with the prevalence rate of enteric HAdV gastroenteritis among all HAdV gastroenteritis cases of 68%, a serotyping study by PCR detected serotype 40 in 46.7% of cases (7/15) and serotype 41 in 53.3% of cases (8/15) with no statistically significant difference between them. The study confirmed that rotavirus and HAdV are prevalent etiological agents of diarrhea in children below the school-age group, highlighting the necessity of the rotavirus vaccine in addition to the obligatory schedule of vaccines in Egypt. Also, it determined that the enteric HAdV gastroenteritis prevalence rate was 68% among all HAdV gastroenteritis.

Role of a schistosoma haematobium specific microRNA as a predictive and prognostic tool for bilharzial bladder cancer in Egypt.

Urinary bladder cancer is a common malignancy in Egypt, thus reliable methodologies are required for screening and early detection. In this study, we analyzed the gene expression of a Schistosoma hematobium specific microRNA "Sha-miR-71a" and mitogen-associated protein kinase-3 (MAPK-3) in the urine samples of 50 bladder cancer patients and 50 patients with benign bilharzial cystitis. Fifty control subjects were also tested. Indirect hemagglutination test (IHA) diagnosed 70% of studied cancer cases as bilharzial associated bladder cancer (BBC), while histopathological examination detected only 18%. Urinary Sha-miR-71a & MAPK-3 revealed enhanced expression in BBC (p-value = 0.001) compared to non-bilharzial bladder cancer (NBBC) cases. Patients with chronic bilharzial cystitis exhibited a significant increase in gene expression compared to those with acute infection (p-value = 0.001). Sha-miR-71a and MAPK-3 showed good sensitivity and specificity in the diagnosis of BBC when analyzed by the receiver operating characteristic (ROC) curve. They were also prognostic regarding malignancy grade. Both biomarkers showed a positive correlation. Our results revealed that IHA is a reliable test in the diagnosis of bilharziasis associated with bladder cancer, and that Sha-miR-71a and MAPK-3 provide non-invasive specific biomarkers to diagnose BBC, as well as a potential role in testing bilharzial patients for risk to develop cancer.

Rapid detection and clinical spectrum of the novel influenza H1N1 strain in a diabetic pediatric population.

OBJECTIVES: H1N1 infection in diabetic patients is of special concern and serious interest since the virus can place individuals, especially children, at great possible risk of subsequently developing type 1 diabetes. This work aims to describe the demographic characteristics, clinical features, and severity of illness of children with type 1 diabetes mellitus (DM), compare the incidence of pandemic H1N1 virus in children with that of the general pediatric population with influenza-like symptoms, and identify the complications of H1N1 virus infection associated with glycemic control. METHODS: The present study included 45 children and adolescents with type 1 diabetes, who were subject to clinical and laboratory investigations. Another 30 healthy adolescents and children with a mean age of 10.43 ± 4.38 years were included as a control group. H1N1 reverse-transcriptase quantitative PCR (RT-Q PCR) was tested for H1N1 virus detection. RESULTS: Diabetic patients positive for (H1N1) showed significantly higher random blood sugar (RBS) levels than diabetic patients negative for (H1N1). Moreover, the H1N1-positive patients had significantly higher hemoglobin (Hb) g/dL, platelet counts, total leukocyte counts (TLCs), and CRP levels. Newly diagnosed patients who were tested positive for (H1N1) and diabetic ketoacidosis (DKA) had significantly higher RBS levels and TLCs than patients who were presented with hyperglycemia. CONCLUSION: RT-PCR is a rapid and specific method for influenza A (H1N1) virus diagnosis. In addition, early administration of oseltamivir no later than 48 hours after the infection is highly recommended in either diabetic or DKA patients suspected of having H1N1.

Molecular study of resistance of Staphylococcus aureus to antiseptic quaternary ammonium compounds.

OBJECTIVES: This study determined the prevalence of qac and smr genes in clinical Staphylococcus aureus isolates from hospital-acquired infections and their susceptibility to quaternary ammonium compounds (QACs) and antibiotics, and correlated the presence of antiseptic resistance genes with antibiotic resistance. METHODS: Susceptibility of 150 non-duplicate clinical S. aureus isolates to antimicrobials and benzalkonium chloride (BAC) was determined by disk diffusion and MIC method, respectively. Resistant strains were analysed by multiplex PCR for the presence of qac and smr genes. RESULTS: Reduced susceptibility to BAC was detected in 30% of isolates (MIC cut-off >8mg/L). QAC resistance genes were detected in 13 isolates with reduced BAC susceptibility. The most frequently detected genes were qacA/B (10 isolates; 22.2%), followed by qacJ (10; 22.2%), smr (8; 17.8%), qacG (8; 17.8%) and qacH (3; 6.7%). There was a strong positive correlation between presence of QAC resistance genes and higher BAC MIC associated with qacA/B, qacJ and smr genes. There was a statistically significant prevalence of antiseptic resistance genes among isolates resistant to cefoxitin, ciprofloxacin, clindamycin, oxacillin, tetracycline and erythromycin. CONCLUSION: This study highlights the prevalence of qac and smr genes in clinical S. aureus isolates with resistance to QACs. There was an association between the presence of antiseptic resistance genes and resistance to different antibiotics, which may be attributed to the presence of both groups of genes on the same plasmid or to selection of resistant strains. More studies are needed on the clinical relevance of the presence of genes controlling resistance to antiseptics.

Role of Hepatitis C Virus (HCV) core antigen in improving blood transfusion safety in high prevalence, resource limited countries, a step forward.

OBJECTIVE: The aim of the present study was to evaluate the efficacy of hepatitis C virus [HCV] core Ag as an alternative affordable test in resource limited countries blood banks. BACKGROUND: Implementing nucleic acid testing in developing countries with low resources is still unaffordable. Egypt has the highest prevalence of hepatitis C in the world and still in need to efficient affordable transfusion program that reduces the window period for the virus before implementing the complex high cost NAT. STUDY DESIGN AND METHODS: HCV core Ag by ELISA in serum, in the presence or absence of anti-HCV antibodies was compared to HCV- RNA by PCR on total number of 1850 first time and repeat donations from Fayoum University Hospital and Badr University Hospital. RESULTS: Among 1850 healthy voluntary donors, 143 donors with anti-HCV antibody positivity, 105 were determined as positive, 38 were negative for HCV core Ag, and 107 were positive for HCV RNA. CONCLUSION: Hepatitis C virus core antigen-ELISA can be a useful alternative in the developing nations and Greater consideration should be given to its implementation as an additional serological test for blood donors in Egypt as the most cost-effective measure for further improvement of transfusion safety.