Cloning of Hepatitis B Virus Surface Gene Region to Escherichia coli

In this study, we cloned HBV s gene to E. coli by using PCR-dependent TA cloning procedure. Our aim was to achieve gene transfer via general principles of recombinant DNA technologies and to use the cloned DNA fragments in future expression studies to obtain antigens to be utilized for diagnostic and immunization procedures

Identification of Beta-Lactamase Enzymes by Isoelectrical Focusing Method in Acinetobacter baumannii

There has been an important resistance problem to various beta-lactam antibiotics in Acinetobacter baumannii strains which cause nosocomial infections. In this study, isoelectric points of beta-lactamase enzymes of 60 strains of A. baumannii isolated from various clinical specimens were investigated by polyacrylamid gel electrophoresis (PAGE). Beta-lactamase enzymes of the isolates were estimated by evaluating their susceptibility to beta-lactam group antibiotics and isoelectric points (pI). The presence of beta-lactamase enzymes was detected in all of the A. baumannii species included in the study. Beta-lactamase band was observed in 50 species by IEF method, but couldn’t be shown in the other 10 isolates. However, the presence of betalactamase which shows spreading type around loading space without band formation was detected in all of the strains. Isoelectric points in 29 of the 50 isolates with band formation were 5.4, 8 were 6.3 and 13 were 6.3 and 5.4. According to antibiotic susceptibility and isoelectric points, 6 phenotypes were observed. We suggest that phenotype 1 was either ACE + TEM-1 or PER-1; phenotype 2 was ACE + CARB-5; phenotype 3 was ACE; phenotype 4 was either an over synthesis of ACE or a new beta-lactamase; phenotype 5 was ACE + TEM-1; phenotype 6 was either ACE + TEM-1 or PER-1 + CARB-5. Every phenotype was divided into subgroups according to their resistance to imipenem, except phenotype 5. As well as beta-lactamases, changes in porins and penicillin binding proteins were considered to be responsible for the resistance of A. baumannii strains to beta-lactam antibiotics

A new line method; A direct test in spinal muscular atrophy screening for DBS

Background: Nucleic acid-based assays provide an opportunity to screen for genetically encoded diseases like spinal muscular atrophy (SMA), before the onset of symptoms. Nowadays, such assays could be easily utilized as high-throughputs in SMA to detect a homozygous deletion of exon 7 of the survival motor neuron 1 gene (SMN1) that is responsible for >95% of SMA patients.Methods: We developed a new line method (NLM) as a direct real time PCR test procedure without nucleic acid extraction in dried blood spots (DBS) to screen for homozygous deletion of exon 7 of the SMN1 gene. Performance of this setup was evaluated on 580 DBS newborn samples and air dried 50 DBS from whole blood including 20 samples for homozygous deletion of the SMN1 gene detected earlier with MLPA.Results: We found all 580 newborn DBS samples as wild type. DBS prepared from 50 whole blood samples also including 20 affected people were correctly identified as homozygous deletions and 30 wild types of exon 7 of SMN1 as before with MLPA. When the MLPA method was taken as the gold standard, the sensitivity and specificity of the NLM test were found 100% for the detection of SMN1 exon 7 homozygous deletion.Conclusion: In the NLM, the total test duration has been reduced to less than 75 min without requiring any extra process such as DNA extraction step and sample plate preparation after the punching step. Thereby, newborn SMA screening with the NLM has gained an environmentally friendly feature with not requiring additional tedious steps.We thank all the newborns, patients and their family members who participated in this study, and we thank the SNP Biotechnology R ; D Ltd. team for their environmental friendly approach and work.We thank all the newborns, patients and their family members who participated in this study, and we thank the SNP Biotechnology Ramp;amp;D Ltd. team for their environmental friendly approach and work

Effect of oral ribavirin treatment on the viral load and disease progression in Crimean-Congo hemorrhagic fever

SummaryObjectivesCrimean-Congo hemorrhagic fever (CCHF) is a lethal hemorrhagic disease. There is currently no specific antiviral therapy for CCHF approved for use in humans. In this study we aimed to investigate the effect of oral ribavirin treatment on the viral load and disease progression in CCHF.MethodsThe study population was composed of patients who had a definitive diagnosis of CCHF by means of clinical presentation plus detection of viral RNA by reverse transcriptase polymerase chain reaction (RT-PCR). Ten patients who received oral ribavirin for 10 days and 40 control patients who received supportive treatment only were included in the study. Ribavirin treatment consisted of oral ribavirin 4g/day for 4 days and then 2.4g/day for 6 days. Viral load and hematological and biochemical laboratory parameters, which were measured daily, were analyzed.ResultsMean age (37.4 vs. 45.5, p=0.285), gender (male 50% vs. 62.5%, p=0.470), days from the appearance of symptoms to admission (4.3 vs.4.4 days, p=0.922), and initial complaints were similar between the ribavirin group and the control group. Upon hospital admission, mean viral load was 8.2×108 copies/ml in the ribavirin group and 8.3×108 copies/ml in the control group (p=0.994). During follow-up, no statistically significant differences were found between the groups with regard to the decrease in viral load, the reduction in alanine aminotransferase and aspartate aminotransferase levels, and the increase in platelet count. The case-fatality rate was 20% (2/10 patients) in the ribavirin group and 15% (6/40 patients) in the control group (p=0.509).ConclusionIn this study, oral ribavirin treatment in CCHF patients did not affect viral load or disease progression

Detection of Crimean-Congo hemorrhagic fever virus genome in saliva and urine

Background: The Crimean-Congo hemorrhagic fever (CCHF) virus is transmitted by tick bites and by contact with the blood or tissues of infected patients and livestock. This study was designed to investigate the genome of CCHF virus in saliva and urine samples of patients with CCHF