Circulating microRNAs as predictors of response to sofosbuvir + daclatasvir + ribavirin in in HCV genotype-4 Egyptian patients.

BACKGROUND: MicroRNAs (miRNAs) play an important role in various diseases, including HCV infection, the aim of the current study was to evaluate the potential use of serum miRNAs as biomarkers for diagnosis, prognosis, and prediction of responses to direct acting antivirals (sofosbuvir + daclatasvir + ribavirin) in HCV-4 patients. METHODS: The serum expression profiles of four liver-associated miRNAs (miRNA-122, 155, 196 and 29) were assessed in 160 HCV-4 patients and 50 healthy controls using real-time PCR prior to therapy. RESULTS: miR-122 and miR-155 showed upregulation in HCV-4 patients compared to healthy controls while miR-196 and miR-29 showed downregulation in HCV-4 patients. ROC curve analyses revealed that the four-studied miRNAs could be valuable biomarkers for predicting response to DAAs with AUC 0.973 for miR-122, 0.878 for miR-155, 0.808 for miR-29 and 0.874 for miR-196 respectively. Univariate logistic regression analysis revealed that miR-196 level is positive predictor for SVR, whereas miR-122,155 levels are negative predictors of response. Multivariate logistic regression analysis revealed that miR-196 is the most significant in predicting response to treatment (p value = 0.011). CONCLUSION: To the best of our knowledge, the current study provided the first clinical evidence of the potential use of circulating miRNAs (miR; 122, 155, 196 and 29) as biomarkers of CHC in HCV-4 patients receiving the new DAA regimen (SOF/DAV + RIB), which is a strong motivator for further studies.

Multidrug-resistant Klebsiella pneumoniae in hospital-acquired infections: Concomitant analysis of antimicrobial resistant strains.

BACKGROUND: Hospital-acquired infections caused by K pneumoniae are difficult to eradicate since K pneumoniae carries resistance genes for many antimicrobials, including carbapenems. The study aimed to determine the prevalence of hospital-acquired infections caused by multiple drug-resistant K pneumoniae and identify carbapenem and fluoroquinolone resistance by phenotypic and genotypic methods amongst hospitalised patients. METHODS: Two hundred and fifty samples from patients with hospital-acquired infections were included. Identification and susceptibility testing for K pneumoniae isolates was performed by standard methods. The detection of carbapenemase resistance (blaKPC , blaVIM-1 and blaOXA-48 ) and plasmid-mediated quinolone resistance (PMQR; qnrA, qnrB and qnrS) genes was performed using PCR assay. RESULTS: Out of 250 samples, 42 (16.8%) were multiple drug-resistant K pneumoniae, and the frequency of K Pneumoniae isolation was higher in urine samples, in the age group (<10 years), in ICU and in patients with longer hospital stay. Twenty-four (57%) of the isolates were resistant to Meropenem, 13 (31%) were resistant to Imipenem and 35 (83.3%) were resistant to Ciprofloxacin. blaOXA-48 gene was detected in 9 (21.4%) of isolates, and blaVIM-1 gene was detected in 6 (14.3%) of isolates. However, no isolate harboured blaKPC gene. PMQR genes were detected in 100% of ciprofloxacin resistant isolates, and qnrS was the dominant. CONCLUSION: Multidrug-resistant K pneumoniae isolates harbouring blaOXA-48, blaVIM-1 and PMQR genes are emerging in hospitals particularly with long hospital stays.

Impact of co-existence of PMQR genes and QRDR mutations on fluoroquinolones resistance in Enterobacteriaceae strains isolated from community and hospital acquired UTIs.

BACKGROUND: Fluoroquinolones are commonly recommended as treatment for urinary tract infections (UTIs). The development of resistance to these agents, particularly in gram-negative microorganisms complicates treatment of infections caused by these organisms. This study aimed to investigate antimicrobial resistance of different Enterobacteriaceae species isolated from hospital- acquired and community-acquired UTIs against fluoroquinolones and correlate its levels with the existing genetic mechanisms of resistance. METHODS: A total of 440 Enterobacteriaceae isolates recovered from UTIs were tested for antimicrobial susceptibility. Plasmid-mediated quinolone resistance (PMQR) genes and mutations in the quinolone resistance-determining regions (QRDRs) of gyrA and parC genes were examined in quinolone-resistant strains. RESULTS: About (32.5%) of isolates were resistant to quinolones and (20.5%) were resistant to fluoroquinolones. All isolates with high and intermediate resistance phenotypes harbored one or more PMQR genes. QnrB was the most frequent gene (62.9%) of resistant isolates. Co-carriage of 2 PMQR genes was detected in isolates (46.9%) with high resistance to ciprofloxacin (CIP) (MICs > 128 µg/mL), while co-carriage of 3 PMQR genes was detected in (6.3%) of resistant isolates (MICs > 512 µg/mL). Carriage of one gene only was detected in intermediate resistance isolates (MICs of CIP = 1.5-2 µg/mL). Neither qnrA nor qnrC genes were detected. The mutation at code 83 of gyrA was the most frequent followed by Ser80-Ile in parC gene, while Asp-87 Asn mutation of gyrA gene was the least, where it was detected only in high resistant E. coli isolates (MIC ≥128 µg/mL). A double mutation in gyrA (Lys154Arg and Ser171Ala) was observed in high FQs resistant isolates (MIC of CIP < 128 µg/mL). CONCLUSION: FQs resistance is caused by interact between PMQR genes and mutations in both gyrA and parC genes while a mutation in one gene only can explain quinolone resistance. Accumulation of PMQR genes and QRDR mutations confers high resistance to FQs.

First detection of vanB phenotype-vanA genotype vancomycin-resistant enterococci in Egypt.

INTRODUCTION: Enterococci have emerged in last two decades as serious hospital acquired pathogens particularly vancomycin resistant strains (VRE). The study aimed to identify the prevalence of enterococcal isolation from hospital infections and colonization as well as determine vancomycin resistance phenotypes and genotypes. METHODS: Sixty enterococcus isolates were isolated from patients, health care workers and hospital environment, identified and tested for antimicrobial susceptibility. Enterococcus species were identified by Real-time PCR and vancomycin resistance was assessed by agar dilution method and Real-time PCR. RESULTS: out of 300 samples (20%) were enterococci (53.3% were E. faecium, 31.7% E. faecalis and 10% other enterococci). Among of them 40/60 (66, 6%) were isolated from infections and 33.3% were isolated from colonization. multiple drug resistance was reported in (100%) of isolates, while (95%) and (45%) of isolates were resistant to vancomycin and ticoplanin respectively. VanA phenotype, vanA genotype was identified in (47.4%) of isolates, while vanB phenotype, vanA genotype was identified in (33.3%) of vancomycin resistant isolates. CONCLUSION: VanB phenotype-vanA genotype was identified in (33.3%) of vancomycin resistant enterococcal isolates. To our knowledge it is the first identified incidence of such strains in Egypt and Africa.