A comparative evaluation of five phenotypic methods for identification of carbapenemase-producing Enterobacteriaceae: a modified carbapenemase detection test.

Rapid detection of carbapenemase-producing Enterobacteriaceae (CPE) is urgently needed to prevent their spread in healthcare settings. Here, we have evaluated the performance of the phenotypic methods for detection of carbapenemase production directly from bacterial cultures. A total of 99 clinical and rectal Enterobacteriaceae isolates were included (81 carrying known carbapenemase-encoding genes and 18 without carbapenemase production). All isolates were subjected to the five phenotypic tests including in-house Carba NP (iCarba NP), modified-Carba NP, E-Test MBL, modified Hodge test (MHT), and commercial combination disk test. Test results were read at different time points for iCarba NP and modified-Carba (1 min, 5 min, 15 min, 1 h and 2 h). The sensitivity and specificity of the iCarba NP were 78.87% and 100%, respectively, whereas those of the modified-Carba NP test were 95.06% and 94.44%, respectively. False-negative results were detected in four OXA-48 isolates with the use of modified-Carba NP, whereas one non-carbapenemase isolate had false-positive results. The sensitivity/specificity was 91.30%/100% and 80.25%/83.33% for the E-Test MBL and MHT, respectively. The sensitivity and specificity of the aminophenylboronic acid synergy test were 100% and 97.94%, respectively, whereas those of the dipicolinic acid synergy test were 82.61% and 96.23%, respectively. Rapid, simple, and reliable methods are needed for laboratory detection of CPE isolates to improve the detection and surveillance of these clinically relevant pathogens in an epidemiological context. We conclude that the modified-Carba NP test can be one of the reliable tests for the prediction of carbapenemase-producing bacteria.IMPORTANCEThe emergence of carbapenem resistance among Gram-negative bacteria is a serious global health threat. Here, we investigate the performance of the five phenotypic assays against carbapenemase-producing and carbapenemase-non-producing Enterobacteriaceae. Accurate and rapid detection of CPE isolates is critically required for clinical management and treatment of infections caused by these organisms. Among the five evaluated phenotypic tests, the mCNP test presented the highest sensitivity (95.06%) and, therefore, can be considered the best test to be used as a screening phenotypic methodology.

Molecular Characterization of Streptococcus Pneumoniae from Patients Diagnosed with Pneumonia: Recommendation for Vaccination Program.

Background: Infections caused by Streptococcus pneumoniae (S. pneumoniae) have remained a significant public health concern worldwide. In developed countries, the highest prevalence of S. pneumonia has been reported among the elderly. The aim of this study was to evaluate the coverage of genotypes in the 13-valent pneumococcal conjugate vaccine (PCV-13) in the Iranian elderly population. Methods: A total of 41 isolates of S. pneumoniae were collected in the current retrospective cross-sectional study. The samples comprised 33 inpatients hospitalized for pneumococcal pneumonia and 8 outpatients. Multiplex polymerase chain reaction assay was performed to categorize the bacteria isolated into specific genotypes. Statistical analyses were performed using SPSS software, and the chi-square test was used to assess the statistical significance in percentages. Results: A total of 68 genotypes were identified in this study, in which 39 isolates (57.3%) were associated with invasive infections. The most common genotypes were 6A/B [8 (19.5%)], 1 [7 (17.5%)], 14 [5 (12.2%)], and 19A [4 (9.75%)], respectively. The coverage rates of PCV-7, PCV-10, and PCV-13 vaccines were 51.17%, 70.7%, and 99.9%, respectively. According to our results, the pneumococcal coverage rate of PCV-7, PCV-10, and PCV-13 vaccine types is estimated to be 51.2%, 70.7%, and 99.9%, respectively. Furthermore, the trend of pneumococcal serotypes included in the PCV-13 was steadily increasing during the study period. Conclusion: It can be concluded that the most circulating pneumococcal serotypes were in accordance with specific serotypes included in the PCV-13 vaccine types. Therefore, including PCV-13 vaccines in immunization programs against pneumococcus in the elderly can effectively reduce the rate of infections.

Virulence genes and antimicrobial resistance pattern in Proteus mirabilis strains isolated from patients attended with urinary infections to Tertiary Hospitals, in Iran.

Background: Proteus mirabilis is a frequent reason for catheter-associated urinary tract infections (UTIs). The aim of this study was to identify virulence genes and antimicrobial resistance patterns in P. mirabilis strains isolated from patients who attended a tertiary hospital in Iran. Methods: In this study, 100 P. mirabilis strains from urine samples were isolated. These isolated strains were identified by biochemical and PCR-based tests, and their antibiotic resistance was profiled through a standard procedure using 14 antibiotics. PCR assays were used to detect virulence-related genes in P. mirabilis strains. The biofilm formation of each P. mirabilis strain was examined. Results: Of the 100 P. mirabilis isolates, 16 (16%) were multidrug-resistant. High resistance was observed against cotrimoxazole (97%), nalidixic acid (93%), cefotaxime (77%), and amoxicillin (62%). Sixty of the 100 isolates showed resistance against extended-spectrum cephalosporins. The prevalence rates of the genes related to the virulence factors in this study were mrpH (100%), ucaA (91%), hpmA (94%), zapA (95%), ptaA (100%), ureG (100%), pmfA (100%), fliC (97%), and mrpA (90%) using PCR method. Strong biofilm formation was observed in 20% (5/25) of the strains isolated from non-catheterized samples and 80% (20/25) of strains isolated from catheterized samples. Conclusions: Resistance to antibiotics and the prevalence of pathogenicity genes are high in Proteus mirabilis strains iolated from UTIs.

Immuno-proteomics analysis between OMV of vaccine and dominant wild type strains of Bordetella pertussis in Iran.

BACKGROUND AND OBJECTIVES: Despite widespread vaccination programs against pertussis, there has been a worldwide resurgence of the disease in recent years. We aimed to investigate protein composition of outer membrane vesicles (OMV) of Bordetella pertussis (Bp) and to evaluate the immunogenicity of OMV antigens both in the vaccine and the dominant wild type strains in Iran. MATERIALS AND METHODS: The OMV were purified from both vaccine and wild type strains. The immunoreactivity of the OMVs was investigated by exposing sera taken from the patients and the vaccinated infants. The protein profiles of OMVs were compared using two-dimensional electrophoresis. The LC-MS/MS was used to analyse and identify differentially expressed protein spots. RESULTS: The two type strains showed differences in their 2D gel protein profile. Further analysis of selected proteins from the dominant Iranian strains using LC-MS/MS demonstrated that the identified proteins fell into different functional categories including (i) metabolism, (ii) membrane transport and secretion system, (iii) biosynthesis and degradation, (iv) adaption, adhesion, pathogenicity, conserved hypothetical and protection responses. Moreover, a number of immunogenic proteins were identified including Bp 2434 (serine protease) and Bp 1616 (putative DNA binding protein) from the vaccine and the wild type strains, respectively which could be considered as potential antigens for an OMV vaccine. CONCLUSION: OMV Bp could be considered as an alternative vaccine against pertussis, containing the bacterium's protein antigens that can confer equal efficacy compared to a whole bacterial cell vaccine with advantages such as less side effects and lower costs than acellular pertussis vaccines.

Molecular detection of six virulence genes in Pseudomonas aeruginosa isolates detected in children with urinary tract infection.

Although a vast majority of Urinary tract infections (UTIs) are caused by E. coli, epidemiological reports have indicated an increasing rate of such infections caused by some other opportunistic organisms including Pseudomonas aeruginosa. Antimicrobial susceptibility and pathogenesis mechanisms of P. aeruginosa are poorly understood. The aim of this study was to detect some virulence factor genes and antimicrobial susceptibility patterns of P. aeruginosa isolates detected in patients with UTI, in children hospital of Tehran, Tehran, Iran. Eighty-four Pseudomonas aeruginosa were isolated. Then, the presence of six virulence genes, in the genome of the isolates was evaluated using PCR amplifications techniques. Finally, antimicrobial susceptibility pattern of the isolates was determined by disk diffusion method. According to the results, lasB was the most prevalent virulence gene that could be detected in the P. aeruginosa isolates (92.9%) used in this study. This was followed by aprA (81.2%), toxA (69.4%), and algD (60%) genes. Two genes, plcH and plcN, were detected in about 38.8% of the isolates. Additionally, Imipenem was found as the most active agent against the P. aeruginosa isolates used in this research. However, Cefotaxime resistance was observed in most of the isolates. Our P. aeruginosa isolates exhibited a great degree of heterogeneity not only in their virulence genes but also in their antimicrobial susceptibility profiles. Imipenem therapies tend to be among the best choices in the management of UTI caused by P. aeruginosa. As a conclusion, assessment of antimicrobial susceptibility pattern and also analyzing the virulence factors can be highly helpful to develop effective treatment strategies against P. aeruginosa urinary infections.

Measurement of interleukin-6 (IL-6) and erythropoietin (EPO) in umbilical cords of preterm infants with intraventricular hemorrhage in two hospitals in Tehran.

BACKGROUND: Intraventricular hemorrhage (IVH) is an important cause of death in premature infants. This study aimed to assess the association of the umbilical cord plasma levels of interleukin-6 (IL-6) and erythropoietin (EPO) with the occurrence and severity of IVH in premature infants. METHODS: Fifty premature newborns of mothers with chorioamnionitis risk factor were selected via nonprobability sampling. The concentration of the cord plasma's IL-6 and erythropoietin were measured by enzyme-linked immunosorbent assay (ELISA) for 3 days. Finally, all samples underwent sonography for the diagnosis of IVH. Results analyzed statistically. RESULTS: Among the samples, 68.98% of them were diagnosed with IVH grade 1. The most severe IVH cases were detected on the second day. The mean and standard deviation of IL-6 level was 74.71 ± 50.53 in the case group and 24.10 ± 46.10 in the control group. There was a correlation between IL-6 levels and IVH (p = 0.0005). The mean and standard deviation of EPO level was 18.38 ± 15.23 in the IVH group and 6.45 ± 13.48 in samples without IVH. A correlation was detected between EPO level and IVH (p = 0.005). CONCLUSION: The concentration of IL-6 and EPO levels of the cord plasma was higher in the premature newborns with IVH.