A simple multiplex PCR for assessing prevalence of extended-spectrum β-lactamases producing Klebsiella pneumoniae in Intensive Care Units of a referral hospital in Shiraz, Iran

OBJECTIVE@#To identify three common genes (blaTEM, blaSHV and blaCTX-M) responsible for ESBL production in Klebsiella pneumoniae (K. pneumoniae)isolated from Intensive Care Units of Namazi Hospital, Shiraz, Iran.@*METHODS@#A total of 60 non-repetitive nosocomial isolates from 60 patients were selected during 2009-2010. The phenotypic identification of ESBL production was confirmed by Double Disk Synergy Test (DDST) according to CLSI guidelines. The ESBL's genotype was then analyzed by multiplex PCR of blaTEM, blaSHV and blaCTX-M genes and DNA sequencing.@*RESULTS@#The primary susceptibility tests of K. pneumoniae showed that among 10 examined antibiotics, the most resistant and susceptible antibiotics identified in this study were ampicillin and imipenem, respectively. The phenotypic determination of ESBL by DDST showed that 60% (n=36) of isolates produced ESBL. Multiplex PCR of genes among K. pneumoniae isolates showed that 39% (n=18) of them have TEM, 39% (n=18) of them have both CTX-M and TEM and 13% (n=8) of them have TEM, SHV, CTX-M.@*CONCLUSIONS@#Our findings reveal the high prevalence (60%) of ESBL producing K. pneumoniae from ICU patients along with a new pattern of blaTEMdistribution differ from other countries.

Prevention and inhibition of TC-1 cell growth in tumor bearing mice by hpv16 e7 protein in fusion with shiga toxin B-subunit from Shigella dysenteriae

For immunotherapy of human papillomavirus [HPV] -16-associated cervical cancers the E7 protein is considered a prime candidate. However it is a poor inducer of cytotoxic T-cell response, when being used as a singular antigen in protein vaccination. Hence, in this study we focused on the utilization of a vaccine delivery system for prevention or treatment of cervical cancer. In this experimental study, we designed and evaluated a novel fusion protein comprising HPV16 E7 antigen fused to Shiga toxin B-subunit [STxB] as both an antigen vector and an adjuvant. Then we designed two preventive and therapeutic tumor models to investigate the prevention and inhibition of TC-1 cell growth in female C57BL/6 mice, respectively. In each model, mice were immunized with the recombinant protein of E7-STxB or E7 without any adjuvant. We demonstrated that prophylactic immunization of E7-STxB protected mice against TC-1 cells. Also in the therapeutic model, E7-STxB inhibited TC-1 tumor growth inlungs. The results were significant when compared with the immunization of E7 singularly. We concluded that immunization with the E7-STxB protein without any adjuvant could generate anti-tumor effect in mice challenged with TC-1 cells. This research verifies the clinical applications and the future prospects of developing HPV16 E7 therapeutic vaccines fused to immunoadjuvants