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Abstract

Background and objectives:

(ESBL) strain is one of the emerging health related problems in the world recently.

Some of the species of the gram-negative bacilli including Klebsiella Pneumonia &

Escherichia Coli are well known ESBL producing among bacteria, and they cause

uncontrollable infections. This Cross-sectional study was designed to asses the

ESBL producing gram negative bacilli among inpatients of Shohada-ye- ashayer

hospital (Khorram Abad).

Extended Spectrum Betalactamase producing

Materials and methods:

methods. ESBL producing gram negative bacilli were screened with MacConkey

Agars containing 4 mg/liter Ceftazidime and confirmed with double disk synergy

method as recommended by national standard laboratory institute.

Samples were processed with routine laboratory

Results:

positive for ESBL.The most isolated species of ESBL are 20 Klebsiella

pneumonia(8.88%), 10 Escherchiia coli(4.44%) and 10 pseudomonas

aeruginosa(4.44%). The most ESBL producing gram-negative bacilli were Isolated

from urine samples (21 cases 39.62%).and Ten cases (18.86%) from bronchoscopy

sterile samples.

Fifty- there cases (23.55%) of 225 total isolated gram negative bacilli are

Conclusion:

frequently isolated from Shohada-ye-Ashaier Hospital. Regarding the high

resistance of these strains against many of the antibiotics and even against

Carbapenems, health- care providers need to plan controlling policies for such

strains.

The Results indicate that ESBL producing gram-negative bacilli are

Key words:

 

Extended Spectrum Betalactamase.

Hospital acquired infection, Escherichia coli, Klebsiella Pneumoniae,

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Background and objectives: HIV-1 Nef and Vpr antigens have been described as suitable candidates for therapeutic HIV vaccine development. The aim of this study was to generate Nef-Vpr fusion gene construct and to clone the construct into pET-23a (+), a prokaryotic expression vector.
Methods: HIV-1 Nef and Vpr genes were PCR-amplified from the pNL4-3 plasmid using specific primers and Pfu DNA polymerase. Results of PCR amplification were visualized by electrophoresis on 0.8% agarose gel. At first, the amplified Nef fragment was cloned into NheI and BamHI restriction sites of pET-23a expression vector. Next, cloning of Vpr gene was performed into BamHI and HindIII restriction sites of the pET-23a-Nef vector. Finally, purity of the recombinant pET-23-Nef-Vpr construct was determined by NanoDrop spectrophotometry.
Results: PCR amplification of Nef and Vpr genes was confirmed by detection of ~ 620 bp and ~ 291 bp bands, respectively. Cloning of the Nef-Vpr construct into the vector was confirmed by detection of a ~ 911 bp fragment following enzymatic digestion with NheI and HindIII and sequencing.
Conclusion: The successful construction of recombinant fusion plasmid encoding a chimeric Nef-Vpr gene was performed in a prokaryotic expression vector for development of HIV-1 recombinant protein vaccine in near future.