[Retrospective evaluation of the alveolar echinococcosis cases between 1980-2010 in Erciyes University Hospital]. / Erciyes Üniversitesi Hastanesinde 1980-2010 Yillari Arasinda Alveoler Ekinokokkozis Saptanan Olgularin Retrospektif Degerlendirilmesi.

OBJECTIVE: Alveolar echinococcosis is an uncommon parasitic disesae confined to the Northern Hemisphere. There is limited data regarding the incidence of the disease in Kayseri. METHODS: Clinicopathologic features of the cases with the diagnosis of alveolar echinococcosis reviewed between 1980-2010. RESULTS: Twenty-nine cases of alveolar echinococcosis were found. There were no significant distribution differences during the study period. 28 of the 29 cases were localised in the liver, whereas one case was localised in the omentum. Sixteen of the 29 cases were male (55%) and 13 were female (45%). The age distribution of these cases varied between 33 and 80. Thirteen cases resided in Kayseri, 2 cases resided in Erzurum, 1 each case resided in Adana, Ardahan, Kars, Nigde, Nevsehir and Yozgat. We could not obtain information from the remaining 8 cases. Abdominal pain was the main symptom in 8 cases, jaundice in 2 cases and fatigue and fever in one case on admission. One case was detected incidentally. All of the cases were diagnosed by histologic examination. CONCLUSION: The data about the alveolar echinococcosis is limited due to its low prevelance. Alveolar echinococcosis cases were detected in Kayseri with a lower incidence than in the East Anatolian region. This report will add data about the incidence of the alveolar echinococcosis.

[Investigation of cytomegalovirus positivity in the peripheral blood samples of risky patients by shell-vial cell culture, antigenemia test and real-time polymerase chain reaction]. / Riskli Hastalarin Periferal Kan Örneklerinde Sitomegalovirus Varliginin "Shell Vial" Hücre Kültürü, Antijenemi Testi ve Gerçek Zamanli Polimeraz Zincir Reaksiyonu ile Arastirilmasi

Cytomegalovirus (CMV) infections in immunocompromised patients and congenital infections in infants have high morbidity and mortality while it may lead to asymptomatic infections in immunocompetent subjects. Serological tests, culture methods, antigenemia tests and molecular methods are applied in the diagnosis of CMV infection. The aim of this study was to investigate the presence of CMV in peripheral blood samples of patients who were at risk for CMV disease by shell vial cell culture, antigenemia test and real-time polymerase chain reaction (RT-PCR) methods. A total of 141 blood specimens obtained from 91 patients (33 female, 58 male) with suspected CMV disease were included to the study. Five of the patients were newborns and the others aged between 17-79 years old were bone morrow (n= 81), kidney (n= 4) and liver (n= 1) transplantation patients. Shell vial (Vircell, Spain) cell culture method was applied for CMV isolation from the samples, while the detection of pp65 antigen in blood leukocytes was investigated by indirect immunofluorescence method (CINAkit Argene, Biosoft, France). The presence of CMV DNA in plasma samples was detected by RT-PCR (CMV QNP 2.0 kit; Fluorion, Iontek, Turkey) method. CMV was found positive in 72 (51%) of 141 samples by shell vial, 82 (58.2%) by antigenemia test and 49 (34.8%) by RT-PCR. Considering cell culture as the gold standard, the sensitivity and specificity of antigenemia test were calculated as 81.9% and 66.6%, respectively; and for PCR those rates were 43% and 73.9%, respectively. In addition DNA sequencing (ABI Prism 310 Genetic Analyzer; Perkin Elmer, USA) was performed for the samples of randomly selected three patients out of 15, who were yielded positive results with cell culture and antigenemia tests but negative CMV DNA by RT-PCR. In this analysis CMV DNA was found positive in three of the samples that were found negative by RT-PCR in spite of CMV isolation and positive antigenemia. DNA sequencing of those samples revealed multiple mutations in the probe binding region (gB) of CMV QNP 2.0 kit. It was concluded that for the detection of CMV viremia and viral load in patients under risk for CMV disease, antigenemia and PCR based methods could be applied, however, negative results obtained by PCR targeting CMV gB gene, should remind the possible presence of mutations in the related site and the results should be confirmed by sequence analysis.

[Investigation of respiratory syncytial virus by three different methods in children with lower respiratory tract infection]. / Alt solunum yolu enfeksiyonu olan çocuklarda solunum sinsityal virus varliginin üç farkli yöntemle arastirilmasi.

Respiratory Syncytial Virus (RSV) is the most important viral agent leading to lower respiratory tract infection in infants and children. The aim of this study was to investigate the presence of RSV by direct immunofluorescence antibody (DFA), cell culture and polymerase chain reaction (PCR) in children with lower respiratory tract infection. Nasotracheal aspirate specimens collected from 80 hospitalized patients aged between 0-24 months and clinically diagnosed as lower respiratory tract infection, during November 2005-May 2006 period, were included to the study. RSV antigen was investigated in clinical specimens by DFA method (Monofluo Bio-Rad, France). Hep-2 culture was used for isolation of RSV. RSV-RNA was investigated by real-time PCR (Fluorion lontek, Turkey) in clinical specimens. RSV was found positive in 26 (32.5%) of 80 samples by DFA and in 17 (21.3%) samples by cell culture. Six specimens were not studied by PCR as sample amounts were not sufficient. Of the 74 samples tested, 20 (27%) were found to be positive by real-time PCR. Fifty-four of the samples were negative by 3 of the methods, while 12 were positive by all of them. DFA and PCR positive 8 samples yielded negative result in cell culture. Five of the 6 samples not investigated by PCR, were positive both in DFA and cell culture while 1 sample was positive only by DFA. Considering cell culture as the gold standard, the sensitivity, specificity, positive and negative predictive values were found as 100%, 85.7%, 65.4% and 100%, respectively, for DFA and 100%, 94.7%, 85% and 100%, respectively, for PCR. As a conclusion for the accurate diagnosis of RSV infections the clinical samples should be collected in the early phase of the disease and inoculated to the cell cultures immediately for viral isolation. If cell culture or PCR facilities are not available for routine diagnosis, DFA method can be used for rapid and cost effective diagnosis of RSV infections.

Comparison of real-time, and qualitative polymerase chain reaction assays in detection of cytomegalovirus DNA in clinical specimens.

OBJECTIVE: To compare the real-time (RT), and qualitative (Q) polymerase chain reaction (PCR) assays for detection of Cytomegalovirus (CMV) DNA. METHODS: The study took place in the Department of Microbiology, Erciyes University, Kayseri, and in Iontek Laboratory, Istanbul, Turkey, from August to December 2006. One hundred and seven clinical specimens from 67 patients were included in the study. Cytomegalovirus DNA was investigated using RT-PCR kit (Fluorion Iontek, Turkey) and Q-PCR kit (Fluorion Iontek, Turkey). Deoxyribonucleic acid sequencing was applied to the samples that yielded discrepant results in both assays. MacNemar's Chi Square test was used for statistical analysis. RESULTS: Of the specimens, 27 were found positive with both assays; 9 with only RT-PCR, and 11 with only Q-PCR assay. Both assays were found negative in 60 of the specimens. There was a good agreement between the 2 assays in 87 (81.3%) of the specimens. There was no statistical significant difference between the assays (p>0.05). Two of the 11 samples that RT-PCR negative Q-PCR positive, and 3 of the 9 samples that RT-PCR positive Q-PCR negative were found to be CMV DNA positive by DNA sequencing. CONCLUSION: A good level of concordance between RT-PCR and Q-PCR assays for CMV DNA detection has been found.