Multilocus sequence typing analysis of Leishmania clinical isolates from cutaneous leishmaniasis patients of Iran.

Cutaneous leishmaniasis (CL) is mainly caused by L. major and L. tropica in Old World and might be represented as typical skin lesion(s) or sometimes as a spectrum of atypical manifestations. We applied multilocus sequence typing (MLST) to explore genetic variations of Leishmania strains isolated from atypical vs. typical CL patients from Iran. A PCR-sequencing was performed for seven housekeeping genes (g6pd, mpi, asat, icd, 6pgd, fh, and trys) and genetic diversity indices and phylogenetic relationships were analyzed. A total of 41 isolates of L. major (28/41) and L. tropica (13/41) from 21 (51.2%) atypical CL and 20 (48.8%) typical CL cases were included. A set of additional sequences of 41 strains of 17 species of Leishmania were retrieved from databases. Different SNP variations were detected and the highest rate of heterozygous sites was found in g6pd and 6pgd genes (6 sites) for L. tropica and in asat and 6pgd genes (7 sites) for L. major strains. All strains were clustered into 58 unique sequence types (STs) including 17 STs related to 41 strains of Leishmania of this study. Concatenated tree clustered all strains in 6 main clades (A to F) including L. major (clade D) and L. tropica (clade B) strains. Two strains of L. major (codes 28 and 42) with highest nucleotide variations were more close to L. tropica and were grouped in Clade B. All of the STs were related in clonal complexes by using eBURST with the prediction of founder genotypes. A high rate of genetic variations and heterozygocity was evident in L. tropica and L. major strains; nevertheless, there was no significant difference in the diversity of Leishmania strains between typical CL and atypical CL groups. This study represents the first successful application of MLST approach to L. tropica and L. major strains in Iran.

Molecular Analysis of Pseudomonas aeruginosa Metallo-Beta-Lactamase: A First Report of an Iranian Referral Pediatric Hospital.

INTRODUCTION: Pseudomonas aerouginosa is an important opportunistic pathogen which causes clinical infections among ill patients. Metallo- Beta- lactamases (MBLs) are important mechanisms of carbapenem (drug of choice) resistance among Pseudomonas aerouginosa isolates. The aim of this study was to determine B- lactamases genes (bla-genes) in P. aerouginosa isolates and to detect percentage of MBLs among P. aerouginosa isolates in different wards. MATERIAL AND METHODS: Clinical isolates of P. aerouginosa in patients hospitalized at Children's Medical Center were collected in two years using a sterile swab. For differentiation and identification of strains the BHI media, Sytrymaid agar and Oxidase test were used and Kirby Baure method antibiotic susceptibility and PCR assay were performed for detection of bla-genes. RESULTS: Based on the study results from a total of 269 isolates of P. aerouginosa, 39 isolates were found to be imipenem resistant. From these isolates, 19 strains of P. aerouginosa isolates were determined to be MBL producers by phenotypic method. All of the Imipenem resistant P. aerouginosa isolates were examined by PCR for the presence of the bla-genes. All MBL- producing isolates carried bla-IMP Genes. And the results of the antibiogram showed the greatest resistance to the Nitrofurantoin, Nalidixic acid and Cotrimoxazole and Cefixime (100%) and resistance to other antibiotics was also significant. CONCLUSION: Considering the prevalence and clinical importance of MBL producing isolates, rapid identification of them and use of the appropriate infection control measures are necessary to prevent further spread of them by these organisms and to help treatment of Infections.