Background: There are insufficient data about the presence of E. albertii as a causative organism in urinary tract infection in pediatric patients. Objective: The present study aimed to detect E. albertii by polymerase chain reaction (PCR) for detection of uidA, mdh, and lysP genes among isolated E.coli from children with urinary tract infection. Methods: The present study was a cross-sectional retrograde study which was carried out on 100 isolates of phenotypically confirmed E.coli detected in urine samples of children suffering from urinary tract infection. The isolates were subjected to molecular identification by PCR for uidA, mdh, and lysP genes. Results: E. albertii was identified by PCR in 7% of the isolates and E.coli was identified in 93% of the isolates. Two mdh and lysP genes were detected for E. albertii and the uidA gene for E. coli. E. albertii isolates had marked resistance to gentamicin (71.4%), followed by resistance to ciprofloxacin (57.1%), meropenem and imipenem (42.9% each) and ESBL activity by double discs method was reported in 57.1% of the isolates. However, none of the isolates had shown resistance to nalidixic acid and only one isolate had resistance to norfloxacin. There was a statistically insignificant difference between resistance to the used antibiotics such as aztreonam (P=0.083), ampicillin/clavulanate (P=0.5), ciprofloxacin (P=0.69), gentamicin (P=0.3) and ceftazidime (P=1.00). Conclusion: The present study highlights the emergence of E. albertii as a pathogen associated with urinary tract infections in children. There is marked antibiotic resistance of this pathogen, especially toward extended spectrum beta-lactams antibiotics. The identification method depends mainly on genetic studies. Further longitudinal studies with large number of patients are required to verify the accurate prevalence of this bacterium.
Background and Aim: Pseudomonas aeruginosa (P. aeruginosa) is an important causative organism of burn infection. Several virulence factors are implicated in P. aeruginosa colonization and invasion, making P. aeruginosa infection's outcome worse. Type III secretion system (T3SS) effector proteins are among these virulence factors. The present study evaluated the frequency of genes encoding T3SS effectors as a virulence determinant in P. aeruginosa isolates from burn patients.Materials and Methods: Wound swabs were collected from burn patients admitted to the Plastic and Reconstructive Surgery Center, Mansoura University, Egypt, and identified by different microbiological testing methods. The modified Kirby Bauer's disc diffusion method was used to test the antibiotic susceptibility of P. aeruginosa isolates against different antibiotics. Prevalence and the presence of exo genes that encode type T3SS proteins (exoS, exoT, exoU, and exoY) in P. aeruginosa isolates were evaluated by the multiplex PCR. Chi-square and Fisher's test were used for statistical analysis.Results: A total of 45 P. aeruginosa isolates were identified from 101 burn patients, including 27 males and 18 females, with a mean age of 15.78±2.65 years old. P. aeruginosa isolates were mostly susceptible to piperacillin/tazobactam and imipenem (73.33 and 62.22%), respectively; while the lowest susceptibility rates were in ceftazidime (4.44%), Tobramycin (4.44%), and ceftriaxone (6.67%). The exoY and exoT genes were detected in 100% of the P. aeruginosa isolates, while 62.22% and 42.22% of clinical isolates harbored exoS and exoU genes, respectively. Conclusion:This study established a correlation between T3SS proteins, particularly exoS and exoU genes and antimicrobial resistance in P. aeruginosa isolates from burn infection.
Resistance to broad spectrum β-lactams mediated by AmpC and metallo beta-lactamases (MBLs) enzymes is a rising problem worldwide. The wide dissemination of Gram negative bacteria harboring these enzymes represents a significant clinical threat during the last decade, which is mainly due to treatment failure and restriction of therapeutic options. This problem should be really estimated in our locality with special emphasis on immunocompromised patients. The aim of this study was to isolate Gram negative bacteria from differrent sites of infection among patients with hematological malignancy, and to examine those isolates for AmpC and MBLs production by phenotypic and genotypic methods. Seventy four Gram negative bacterial strains were isolated from 387 clinical samples collected from different infection sites. Those isolates were screened for the presence of AmpC and MBLs by modified three dimensional test and Imipenem-EDTA combined disc test, respectively. Multiplex PCR was done as a confirmatory step for detection of AmpC and MBLs production by these isolates. Pseudomonas aeroginosa was the most common isolated Gram negative strain that was found to be positive for AmpC and MBL production. DHA gene was the most frequently detected AmpC β-lactamase gene, whereas VIM was the only detected MBL gene among the Gram negative bacterial isolates by multiplex PCR. The strong association found between AmpC production and MBL gene carriage is alarming which necessitate continuous surveillance of such resistance mechanisms among the Gram negative bacteria, especially in patients with hematological malignancy.
Mannose binding lectin (MBL) is an important component of innate immunity particularly in neonates whose adaptive immunity is not fully developed. Polymorphism in MBL2 gene promoter and exon1 determines MBL serum level and function. The aim of this study was to investigate the frequency of different MBL2 genotypes in neonatal sepsis among patients of neonatal intensive care unit (NICU). Two hundred and forty-five neonates were enrolled in this study (127 infected and 118 uninfected controls). Multiplex PCR and double amplification refractory mutation system (dARMS) were used for typing of MBL2 exon1 and promoter respectively. Klebsiella species were the most frequently isolated organisms (22.8%). There is no statistical significance difference in the distribution of different expression genotypes between infected group and controls (P = 0.11). However, prevalence of low MBL2 expression genotypes (XA/O and O/O) was higher in infected patients compared to control group (patients 25.2% and controls 15.3%). Low and medium MBL2 expression genotypes were mostly associated with Gram-negative bacterial infections (18.9% and 22.8%) respectively. A statistically significant association of Gram-negative bacterial infections with low MBL2 expression genotypes was found (P = 0.02). Higher frequency of AB and BB genotypes was observed (31.5% and 7.9%) in patients group compared to control, but without statistical significant difference.
Background: Shigella is one of the most serious pathogens associated with bloody diarrhea in children. The empiric antibiotic therapy of enteric illness with blood streaked stool leads to emergence of multi drug resistant (MDR) Shigella. The condition gets exacerbated by presence of integrons that facilitate the horizontal spread. Virulence genes associated with MDR Shigella modulate the patient outcome, particularly in children. Objectives: The present study was aiming at isolation of MDR Shigella from children with diarrheal sickness and characterization of those isolates as regarding presence of class 1 integrase and other virulence genes. Methods: Four hundred and ninety patients under the age of five suffering from diarrheal illness were examined for presence of Shigella in their stool specimens. MDR Shigella was determined using the antibiotic susceptibility testing by disc diffusion method; those isolates were tested for presence of class 1 integrase by PCR. Multiplex PCR assay was used to determine the presence of virulence genes, virA, ial, sen, set1A, set1B, sat, ipaBCD, ipaH and stx in the MDR Shigella isolates. Results: The isolation rate of Shigella from pediatric patients was 5.3%. Most of the isolated Shigella (57.7%) were from infants between 12 and 23 month. 73.1% of the identified Shigella were MDR. intI1 gene was present in 78.9% of MDR isolates. Muliplex PCR revealed that ipaH and ipaBCD, virA, sat, ial, set1A and set1B, sen were detected in 94.7%, 78.9%, 73.7%, 68.4%, 42.1%, 36.8% of the MDR Shigella isolates respectively. Conclusion: The MDR isolates represented a considerable percentage of Shigella detected in pediatric patients. Presence of intI1 gene in most of MDR Shigella reflects the higher possibility of resistant strains spread. Existence of a variety of virulence genes in those isolates is an important indicator of serious disease outcome.
Early diagnosis of urinary tract infection (UTI) in pediatric patients is a problem due to the absence of specific symptoms and difficulty in obtaining the proper urine sample. These difficulties increase the need for effective biomarker for UTI diagnosis in young patients. Neutrophil gelatinase-associated lipocalin (NGAL) is proved to be a marker for UTI diagnosis. YKL-40 is a glycosyl hydrolase which is produced locally at the sites of inflammation. The objective of our study is to assess the value of YKL-40 as a possible marker for UTI diagnosis in febrile children and to compare its value versus NGAL. Urine culture was used as a gold standard for UTI diagnosis. The study enrolled three groups; febrile children with positive urine culture, febrile children with negative urine culture, and controls without fever and with negative urine culture. Each group included 50 children from patients attended outpatients clinics department of Mansoura University Children Hospital. For each patient enrolled in the study, complete blood count, C-reactive protein, serum creatinine, urine creatinine, routine urine analysis and urine culture were assessed. Using ELISA test, urine values of NGAL (uNGAL) and YKL-40 (uYKL-40) were measured and normalized to urine creatinine (uNGAL/uCr) and (uYKL40/uCr) respectively. The values of uNGAL, uYKL-40, uNGAL/Cr and uYKL-40/Cr were significantly higher in febrile UTI group. The receiver operating curve (ROC) show the optimum cut off value for urine YKL-40 (171.5 pg) with 84% senstivity, 82% specificity and the area under the ROC curve (AUC) 0.95. The optimum cutoff value of uYKL-40/Cr was (159.2 pg/mg) with 72% sensitivity, 71% specificity and 0.81 AUC. Higher sensitivity and specificity of uYKL-40 and uYKL40/Cr compared to uNGAL and uN-GAL/Cr respectively were observed. In conclusion, the diagnostic value of uYKL-40 is superior to that of uNGAL. Urine YKL-40 could be a good marker for diagnosis of UTI in febrile pediatric patients.
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