Serdar Tuncer

PCR detected hepatitis C virus genome in the brain of a case with progressive encephalomyelitis with rigidity.

A case of progressive encephalomyelitis with rigidity (PEWR) associated with hepatitis C virus (HCV) is reported. A 58 year-old woman presented with a clinical picture of progressive quadriparesis, sensory loss, sphincter dysfunction, painful muscle spasms in the upper and lower limbs and continuous muscle unit activity in electromyography. She developed hepatitis, pancreatitis and HCV-RNA was detected in the plasma by reverse transcription-polymerase chain reaction (RT-PCR). Postmortem histopathological examination showed encephalomyelitis with perivascular lymphocyte cuffing, infiltration and neuronal loss mainly affecting the brainstem and cervical spinal cord. The RT-PCR analysis of the postmortem brain, brainstem, liver, pancreas, plasma and CSF samples revealed the presence of HCV genome in all specimens except CSF. Clinical features, postmortem histopathology and PCR results and the possible etiopathogenesis of PEWR are briefly discussed.

PCR on disseminated tuberculosis in bone marrow and liver biopsy specimens : Correlation to histopathological and clinical diagnosis

Disseminated tuberculosis with negative pulmonary findings is a diagnostic problem. Histopathological studies of bone marrow (BM) and liver (LV) biopsies are the most reliable methods for diagnosis in such cases; however, their sensitivity is limited. In this retrospective study, 41 BM and 7 LV paraffin-embedded biopsy specimens from clinically (clinical response to antituberculous treatment after 6 months follow-up) and/or histopathologically diagnosed tuberculosis were analysed for the detection of Mycobacterium tuberculosis DNA by polymerase chain reaction (PCR). Two different primer sets, one based on the repeated IS6110 sequence of M. tuberculosis and the other based on the mtp40 gene region, were used for amplification. Histopathological and PCR studies were positive for M. tuberculosis in 12/41, and 30/41 in BM and 4/7, and 6/7 in LV biopsy specimens, respectively. As the control group, 17 BM biopsy specimens obtained from patients with a positive Mantoux skin test but no active tuberculosis were analysed. One BM biopsy out of 17 control cases was positive with PCR while none was consistent with TB histopathologically. In conclusion, PCR might be applicable and more reliable than histopathological studies for detection of tuberculosis in BM and LV biopsy specimens.

Human papillomavirus associated with bladder carcinoma? Analysis by polymerase chain reaction

Background: The aim of the present study was to assess the possible etiologic role of human papillomaviruses (HPV) in bladder tumors.

Comparison of cytochemical staining, immunofluorescence and PCR for diagnosis of pneumocystis carinii on sputum samples

Detection of P. carinii has increased with the use of polymerase chain reaction (PCR), particularly in sputum samples. In this study, sputum samples obtained from 30 immunosuppressed patients with respiratory symptoms (12 HIV-infected) were tested by standard cytochemical staining (Giemsa and methenamine silver), immunofluorescence (IF) staining and PCR for detection of P. carinii and the results were compared. Pneumocystis carinii was detected in 4, 8 and 13 sputum samples by cytological staining, IF test and PCR, respectively. Specific amplification bands were obtained in all sputum samples that were positive by both other tests. All tests gave negative results in sputum samples obtained from 5 HIV-infected asymptomatic patients and 22 non-immunosuppressed tuberculosis patients. Our observations suggest that PCR results were well correlated with P. carinii pneumonia (PCP), especially in non-HIV-infected patients. However, PCR positivity obtained in HIV-infected patients could be misleading in the diagnosis of PCP without careful clinical evaluation. Positive results obtained by Giemsa staining or IF test confirm diagnosis of PCP authoritatively. As a result, we suggest testing sputum samples by both PCR and IF techniques for detection of P. carinii.

DETECTION OF BACILLUS CALMETTE-GUERIN IN THE BLOOD BY THE POLYMERASE CHAIN REACTION METHOD OF TREATED BLADDER CANCER PATIENTS

PURPOSE Following intravesical bacillus Calmette-Guerin (BCG) instillation, we attempted to detect BCG in the blood using the polymerase chain reaction (PCR) method and correlate these findings with the occurrence of major complications due to this treatment. MATERIALS AND METHODS Intravesical BCG immunotherapy was given to 22 consecutive patients with superficial bladder tumors. In 2 patients the BCG instillation had to be discontinued due to serious side effects of therapy. Blood samples (252 aliquots) were obtained from 126 BCG courses in 22 cases, and 2 additional samples (4 aliquots) were obtained from 1 patient 1 and 3 months after cessation of therapy. All blood samples were analyzed by the PCR technique for detection of deoxyribonucleic acid tuberculosis Mycobacterium tuberculosis. RESULTS Of the 126 blood samples 9 (7.1%) were PCR positive for M. tuberculosis. These 9 positive samples belonged to 3 patients, all of whom were among those 4 patients who had major clinical side effects. CONCLUSIONS We demonstrated that rapid and sensitive detection of mycobacteremia by PCR correlated with the clinical course of these patients. We also demonstrated that PCR can be used to monitor BCG in the blood after antituberculous therapy. The early, fast and accurate diagnosis of BCG in the blood by PCR may alter the serious clinical course of these patients by initiation of specific treatment early. However, further extensive studies are needed to validate these results.

Prevalence of Helicobacter pylori in symptomatic patients and detection of clarithromycin resistance using melting curve analysis.

UNLABELLED Abstract. BACKGROUND Clarithromycin is often a component of combination therapies for Helicobacter pylori eradication; however, increases in resistance rates have decreased the success of the treatment. OBJECTIVE This study was designed to determine the prevalence of H pylori infection in symptomatic patients and to detect clarithromycin resistance rates using melting curve analysis. METHODS Patients scheduled for upper endoscopy at the Endoscopy Unit of the Department of Gastroenterology, Duzce University, Medical Faculty Hospital, Konuralp/Duzce, Turkey, were assessed for enrollment in the study. Two pairs of gastric biopsy specimens (antrum and corpus) were obtained from each study patient. Histopathologic examination, rapid urease test, culture, and polymerase chain reaction (PCR) of the specimens were used to identify H pylori infection. Clarithromycin resistance was detected using melting curve analysis. RESULTS Seventy-five patients (41 women, 34 men; mean [SD]age, 42.6 [14.5] years [range, 17-70 years]) were included in the study. Using histopathology and rapid urease test, H pylori was detected in 40 (53.3%) of the 75 specimens. H pylori was detected using PCR in 40 (53.3%) specimens and by culture in 10 (13.3%) specimens. The specificity and sensitivity of PCR and culture were interpreted by comparing them with the results of histopathologic examination and urease tests. The specificity and sensitivity of PCR were 68.6% and 72.5%, respectively, and the specificity and sensitivity of culture were 97.1% and 22.5%, respectively. Of the 40 isolates, 21 (52.5%) were susceptible to clarithromycin, 12 (30.0%) were resistant, and a mixed susceptibility pattern was detected in 7 (17.5%) specimens. H pylori isolates from 19 (79.2%) of the 24 patients who had formerly used clarithromycin showed clarithromycin resistance. CONCLUSIONS The prevalence of H pylori infection was 53.3% for the symptomatic patients in this study, and 47.5% of the isolates showed clarithromycin resistance using melting curve analysis. The PCR-based system used in this study was accurate for the detection of H pylori infection as well as clarithromycin susceptibility testing directly in biopsy specimens.