Comparison of manual and automated nucleic acid isolation methods for HBV-DNA and HCV-RNA assays.

In the diagnosis and monitoring of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections, it is important to use methods that can provide rapid and reliable results. The present study aimed to compare the automated and manual extraction methods during the nucleic acid isolation phase for HBV-DNA and HCV-RNA assays. The study included 93 serum samples, 49 of which were for the HBV-DNA assay and 44 for the HCV-RNA assay. DNA and RNA isolation from the samples was performed manually with a "QIAmpMin Elute Kit" (Qiagen, Germany) and the automated isolation system, NucliSens easyMAG (BioMérieux, France). All the extraction products were amplified using the iCycler device (Bio-Rad, USA). With both methods, compliance was found in 21 (42.8%) samples in the HBV-DNA assay; nine (18.3%) samples had a higher amount of viral nucleic acid with the manual method, whereas 19 samples (38.7%) were found to have a higher amount of nucleic acid with the automated system. For the HCV-RNA assay, total compliance was found in 31 (70.4%) samples; 12 (27.2%) samples had a higher amount of viral nucleic acid with the manual method whereas one sample (2.2%) was found to have a higher amount of nucleic acid with the automated system. It was concluded that the NucliSens easyMAG automated isolation system can be used with confidence for nucleic acid extraction due to its higher sensitivity, providing results in a shorter time, and assured standardization.

Invasive pneumococci before the introduction of pneumococcal conjugate vaccine in Turkey: antimicrobial susceptibility, serotype distribution, and molecular identification of macrolide resistance.

This study evaluates the antimicrobial susceptibilities and serotype distributions of invasive Streptococcus pneumoniae (SP) isolates identified in a Turkish hospital before the introduction of the 7-valent pneumococcal conjugate vaccine (PCV7). The susceptibilities of all isolates were determined by evaluating six antibiotics: penicillin (PEN), ceftriaxone (CRO), levofloxacin (LEV), erythromycin (ERY), clindamycin (CD), and vancomycin (VAN). Serotyping and amplification of macrolide resistance genes were performed. Sixteen (50%) and four (2%) isolates were resistant to PEN and LEV, respectively. No isolates demonstrated VAN resistance. Intermediate resistance to CRO was found in 4% of all invasive isolates. Twenty-three (12.6%) isolates were resistant to ERY. Four (2%) invasive SP isolates demonstrated multidrug resistance. Serogroups 3, 5, 6, 8, 9, and 23 were the most common in both age groups. The potential coverage rates of PCV7 and PCV13 were 44.1 and 66.1% in children and 39.8 and 71.5% in adults, respectively. Continuous surveillance of antimicrobial resistance is required.

Matrix Metalloproteinases 2 and 9 Polymorphism in Patients With Myeloproliferative Diseases: A STROBE-Compliant Observational Study.

Chronic myeloproliferative disorders such as polycythemia vera (PV), essential thrombocytosis (ET), and idiopathic myelofibrosis arise from clonal proliferation of neoplastic stem cells in the bone marrow. Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that have potential to degrade all types of extracellular matrix (ECM) and also play a role in remodeling of the ECM. It is known that MMPs play a role in bone marrow remodeling.The primary goal of our study is to explore the relationship between chronic myeloproliferative diseases and some of MMP gene polymorphisms. The demonstration of a relationship will help to understand whether these polymorphisms may be a potential early diagnosis marker of the diseases.Patients were selected from outpatient clinics of Turgut Ozal University Hospital, Ankara, Turkey, between December 2010 and May 2011. Twenty-eight patients that previously diagnosed and followed-up with PV, 17 with secondary polycythemia (SP), and 12 with ET were enrolled in the study, along with a control group of 22 healthy people.DNA was isolated from peripheral blood. Using polymerase chain reaction-restriction fragment length polymorphism method, MMP2 and MMP9 gene polymorphisms were analyzed with agarose gel electrophoresis. There was a statistically significant difference between the study groups and the control group in terms of Gln279Arg polymorphisms rates of MMP9. The highest MMP9 Gln279Arg polymorphism rate was observed in the ET group. But nobody from the control group had polymorphic MMP9. There was no statistically significant difference between the groups in terms of MMP2-735 C > T polymorphism rates.In conclusion, MMP9 gene Gln279Arg polymorphism was associated with ET, SP, and PV diseases. Hence, we believe that these gene polymorphisms may play a role in the mechanism of bone marrow fibrosis and may be a factor that increases the risk of thrombosis. Illumination of the molecular basis of the relationship between MMP-thrombosis and MMP-fibrosis provides a better understanding of the pathophysiology of PV and ET diseases and will allow new approaches to diagnosis and treatment.

Antimicrobial Susceptibilities of Clinical Acinetobacter baumannii Isolates With Different Genotypes.

BACKGROUND: The treatment of Acinetobacter baumannii infections is difficult. Carbapenems, sulbactam, and colistin are the most effective antibiotics. OBJECTIVES: The aim of this study was to evaluate the susceptibilities of genotypically different A. baumannii isolates to sulbactam, amikacin, netilmicin, meropenem, tigecycline and colistin. PATIENTS AND METHODS: Isolates from various clinical samples of patients with hospital-acquired infections that were identified by the VITEK 2 Compact system in our hospital's microbiology laboratory between January 2010 and March 2012 were included in the study. To determine genetic relatedness of the isolates, the rep-PCR method was used. The broth microdilution method was used for amikacin, netilmicin, meropenem and colistin, while E-test was used for sulbactam and tigecycline. RESULTS: Among the 300 isolates, 30 were found to be genotypically different and were evaluated in terms of their antimicrobial susceptibilities. All isolates were susceptible to colistin. The susceptibility rates were 66.6%, 50%, 36.6%, 30%, and 10% for netilmicin, tigecycline, sulbactam, amikacin, and meropenem, respectively. For carbapenem resistant isolates, the susceptibility rates were 66.6%, 51.8%, 33.3%, and 25.9% for netilmicin, tigecycline, sulbactam, and amikacin, respectively. The sulbactam minimum inhibitory concentration (MIC) 50 and MIC 90 were 8 µg/mL and 12 µg/mL, respectively. CONCLUSIONS: In this study, it was concluded that determining the cut-off value for MIC breakpoints for sulbactam alone has a critical impact on the susceptibility results.

Plasmodium falciparum malaria case originating from Uganda.

Malaria is the fifth infection leading to death in the world. Plasmodium species is the malarial parasite that infects human cells. The five species of the human Plasmodium parasites are P. falciparum, P. vivax, P. ovale, P. malariae and P. knowlesi. Recently, the World Health Organization reported that Uganda has the world's highest malaria incidence, with a rate of 478 cases per 1000 population per year. In this article, a patient who had specific clinical signs and symptoms of malaria after work-related travel to Uganda has been evaluated. The major clinical findings of the patient were chills and fever. After examination of thin and thick blood smears prepared from the peripheral blood of the patient, P. falciparum parasites were observed. Cerebral malaria was suspected as the patient's consciousness, orientation and cooperation had deteriorated. No Plasmodium was seen in control blood smears after treatment.