Evaluation of two colorimetric microplate microtiter tests for susceptibility testing of clinical isolates of Mycobacterium tuberculosis to first-line antituberculosis drugs.

Background: Drug-resistance tuberculosis (TB) is one of the most important global public health problems. Accurate and rapid drug-susceptibility testing is critical for the effective treatment of TB patients. Various colorimetric methods are used for anti-TB drug-susceptibility testing (DST) and minimum inhibitory concentration (MIC) determination. This study was conducted to evaluate the resazurin microtiter assay (REMA) and malachite green decolorization assay (MGDA). Methods: A total of 65 Mycobacterium tuberculosis strains isolated from patients with suspected TB using REMA and malachite green microtiter assay methods were tested against streptomycin (SM), isoniazid (INH), rifampicin (RIF), and ethambutol (ETB). The Mycobacterial Growth Indicator Tube 960 DST method was accepted as the gold standard in the evaluation of test results. Results: The sensitivity of REMA and MGDA tests was found to be 87.5% and 62.5% for INH, respectively. RIF and SM sensitivity for both tests was 100%. While ETB sensitivity was 81.8 for the REMA test, this rate was 60% for the MGDA test. Specificity of both tests varied between 92.5% and 98.2% according to the drug types. Conclusion: REMA and MGDA are a simple, rapid, and low cost. They can be used as an alternative test for drug-susceptibility testing and MIC determination. Extensive studies and standardization are needed for the routine use of both tests.

[Evaluation of second-line antituberculosis drug susceptibilities of multidrug-resistant Mycobacterium tuberculosis complex isolates by E-test method]. / Çok ilaca dirençli Mycobacterium tuberculosis kompleks izolatlarinda ikinci basamak antitüberküloz ilaç duyarliliklarinin E-test yöntemi ile degerlendirilmesi.

Multidrug-resistant (MDR) tuberculosis (TB) constitutes a restricting factor for the effective treatment of TB worldwide. Early diagnosis and appropriate treatment of patients are the most effective strategy in the control of MDR-TB. Therefore, knowledge of drug resistance patterns of the MDR-TB clinical isolates are necessary in planning of an appropriate treatment regimen for the patient. The aims of this study were to detect the susceptibilities of MDR-TB isolates to second-line anti-TB drugs by E-test method, and to compare their results with Löwenstein-Jensen (LJ) proportion method. A total of 122 MDR (resistant to isoniazid and rifampicin) Mycobacterium tuberculosis complex (MTC) strains isolated from samples of patients with pulmonary TB were included in the study. The isolates were identified by conventional methods and first-line anti-TB drug susceptibility testing was performed by the proportion method using LJ medium. The susceptibilities of the isolates to second-line anti-TB drugs [kanamycin (KN), ofloxacin (OFL), ethionamid (ETN), linezolid (LIN)] were tested by proportion method on LJ medium and E-test method on Middlebrook 7H11 medium. For this purpose, E-test strips (bioMerieux, Fransa) of KN (0.016-256 mg/ml), OFL (0.02-32 mg/ml), ETN (0.016-256 mg/ml), and LIN (0.016-256 mg/ml) were used. The susceptibility tests were evaluated in 5., 7., and 10. days after application of the E-test strips, and proportion method on LJ medium was evaluated 28 days later. Second line-anti-TB drug susceptibility results were obtained in 5 to 10 days by E-test. Of the MDR MTC strains 98% (119/122) were susceptible to KN, OFL and LIN, while 2% (3/122) of the strains were resistant to KN and ETN. The correlation between E-test and LJ proportion method was estimated as 96% for KN and ETN, 98% for OFL, and 100% for LIN. When compared with LJ proportion method, the specificity of E-test in the detection of susceptibility to KN, OFL, ETN and LIN were 60%, 38%, 60%, and 100%, respectively, while the sensitivity was 100% for all drugs. Our results indicated that E-test method exhibited high sensitivity and specificity (100%) for LIN, so it may be used alone in susceptibility testing for this drug, however since the specificity is low (38%) for OFL it should be used together with the proportion method. In conclusion, E-test method might contribute for initiation of an early and effective anti-TB drug treatment and control of infection by rapid diagnosis in MDR-TB cases.

Clinical importance of serum neopterin level in patients with pulmonary tuberculosis.

OBJECTIVE: Neopterin is a sensitive marker for cell-mediated immune response. Because of this, the neopterin levels of body fluids show cell-mediated immune response in different infectious diseases which involve T cells and macrophages. The aim of this study was to determine the clinical importance of neopterin levels in patients with tuberculosis and compare with those levels of healthy subjects. METHODS: Seventy patients with tuberculosis (46 newly diagnosed cases, 15 relapse cases, and 9 multidrug-resistant tuberculosis cases) and 18 healthy adult individuals were included in the study. Neopterin concentrations were measured by the ELISA method according to the protocol of the manufacturer. Chi-square test was used in statistical analysis; p⩽0.05 was considered statistically significant. RESULTS: Serum mean neopterin levels were 23.74±21.8nmol/L (median: 18.3) in newly diagnosed patients with pulmonary tuberculosis; 28.69±21.2nmol/L (median: 21.2) in relapse patients and 31.28±14nmol/L (median: 25.4) in multidrug-resistant tuberculosis cases, respectively. Serum mean neopterin levels were 4.03±5.12nmol/L (median: 5.1) in healthy subjects. The serum neopterin levels were found to be significantly higher in patients with tuberculosis than the control group. There was a statistically significant correlation between neopterin positivity (neopterin level ⩾10nmol/L was accepted to be positive) and clinical symptoms of hemoptysis and weight loss. Besides statistically significant correlations between neopterin positivity and hemoglobin level, sedimentation rate, mean leukocyte count and radiological involvement (localized or diffuse) were determined. CONCLUSION: Serum neopterin levels can be used as a helper laboratory finding for the diagnosis of patients with tuberculosis. For this aim, further controlled studies are needed.

[Evaluation of the distribution of non-tuberculous mycobacteria strains isolated in National Tuberculosis Reference Laboratory in 2009-2010, Turkey]. / Ulusal Tüberküloz Referans Laboratuvarinda 2009-2010 Yillarinda Tespit Edilen Tüberküloz Disi Mikobakterilerin Dagilimlarinin Irdelenmesi.

Non-tuberculous mycobacteria (NTM) are commonly encountered environmental bacteria, and most of them are associated with lung diseases. Diagnosis of infections caused by NTM is based on clinical, radiological and microbiological findings. The aim of this study was to investigate the distribution of non-tuberculous mycobacterial species isolated from clinical specimens as etiologic agents. The NTM strains isolated from clinical specimens in National Tuberculosis Reference Laboratory (NTRL), together with the strains that were sent to NTRL for the advanced identification of non-tuberculous mycobacterial species that have clinical or microbiological significance, were analysed retrospectively. The strains belonged to January 2009 - December 2010 period. If the same NTM type was isolated more than once in the clinical specimens of a patient, then it was defined microbiologically as a causative agent. Identification of mycobacteria species was performed by using a commercial line-probe assay (GenoType Mycobacterium CM/AS; Hain Lifescience, Germany). In our study, pulmonary and non-pulmonary samples obtained from 206 patients yielded mycobacterial growth in their cultures, and of them 24 (11.7%) were identified as NTM. On the other hand, 51 of the 101 samples sent to NTRL for identification were confirmed as NTM. Of the patients who were found to be infected with NTM (n= 75), 59 (78.7%) were male and the mean age was 50.9 ± 18.8 years. The most frequently identified NTM species was M.fortuitum (33.3%, n= 25), followed by M.abscessus (18.7%, n= 14), M.gordonae (10.7%, n= 8) and M.avium (%8; n= 6). The other types of NTM species identified in our laboratory were M.chelonae (n= 3), M.intracellulare (n= 3), M.kansasii (n= 3), M.peregrinum (n= 2), M.scrofulaceum (n= 2), M.szulgai (n= 2), M.celatum (n= 1), M.haemophilum (n= 1), M.smegmatis (n= 1) and M.xenopi (n= 1). Rapidly growing NTM species (M.fortuitum and M.abscessus) were the most frequent (52%) species isolated in our laboratory as the cause of non-tuberculous mycobacterial infection. Interestingly, the majority of M.fortuitum isolates (n= 21) which was the most common species identified in our laboratory, were those received from the peripheral laboratories. The most common species identified in our laboratory were rapidly growing NTM, however the countrywide distribution of the NTM species was found different than previously reported. In conclusion, further investigation of the non-tuberculous mycobacteria profile in adjunct with epidemiological data seems to be essential in our country.

[Comparison and evaluation of Lowenstein-Jensen medium and 2% Ogawa medium for the diagnosis of tuberculosis]. / Tüberkülozun Tanisinda %2 Ogawa Besiyerinin Löwenstein-Jensen Besiyeri ile Karsilastirilmasi ve Degerlendirilmesi.

Accurate diagnosis of tuberculosis is based on the detection of the bacilli in the clinical specimen. The growth of mycobacteria in laboratory media is regarded as the gold standard for diagnosis and use of two different cultivation media, one of them being a liquid one, is recommended. In this study, the diagnostic values of egg-based Lowenstein-Jensen (LJ) medium used extensively all over the world and Ogawa (2%) medium were compared. A total of 7912 pulmonary and extrapulmonary clinical samples which belonged to 3311 patients with suspected tuberculosis, were enrolled in the National Tuberculosis Reference Laboratory in Refik Saydam National Public Health Agency during 2.5 years (January 2006- June 2008). The samples were processed by modified Petroff method (4% NaOH) for homogenization-decontamination-concentration on the same day and inoculated on two Ogawa and two LJ media. The cultures were incubated at 35-37°C for eight weeks and were controlled on the third day of incubation in terms of contamination. If no contamination were detected, the cultures were incubated for a total of eight weeks with regular weekly controls. The colonies detected in culture media were identified by biochemical tests, including paranitrobenzoic acid (PNB), niacin accumulation and nitrate reduction tests. Those identified as Mycobacterium tuberculosis complex were reported as positive. The rates of contamination was 3.1% for Ogawa medium and 4% for LJ medium. Culture results were found positive for 248 patients (7.5%). While 210 of these were positive by both of the media, 20 (8.1%) patients were detected positive only by LJ and 18 (7.3%) only by Ogawa medium. The sensitivity of LJ medium was 92.7% (230/248) and of Ogawa medium was 91.9% (228/248). When LJ medium was taken as the reference method, the sensitivity, specificity, positive and negative predictive values of Ogawa medium were 91.3%, 99.4%, 92.1% and 99.4%, respectively. It was concluded that, when low price and low contamination rates were taken into consideration, Ogawa medium, used together with a liquid medium + LJ medium, would increase the yield of mycobacteria from single samples (cerebrospinal fluid, biopsy, pus, etc.) sent to the laboratories.

Distribution of spoligotyping defined genotypic lineages among drug-resistant Mycobacterium tuberculosis complex clinical isolates in Ankara, Turkey.

BACKGROUND: Investigation of genetic heterogeneity and spoligotype-defined lineages of drug-resistant Mycobacterium tuberculosis clinical isolates collected during a three-year period in two university hospitals and National Tuberculosis Reference and Research Laboratory in Ankara, Turkey. METHODS AND FINDINGS: A total of 95 drug-resistant M. tuberculosis isolates collected from three different centers were included in this study. Susceptibility testing of the isolates to four major antituberculous drugs was performed using proportion method on Löwenstein-Jensen medium and BACTEC 460-TB system. All clinical isolates were typed by using spoligotyping and IS6110-restriction fragment length polymorphism (RFLP) methods. Seventy-three of the 95 (76.8%) drug resistant M. tuberculosis isolates were isoniazid-resistant, 45 (47.4%) were rifampicin-resistant, 32 (33.7%) were streptomycin-resistant and 31 (32.6%) were ethambutol-resistant. The proportion of multidrug-resistant isolates (MDR) was 42.1%. By using spoligotyping, 35 distinct patterns were observed; 75 clinical isolates were grouped in 15 clusters (clustering rate of 79%) and 20 isolates displayed unique patterns. Five of these 20 unique patterns corresponded to orphan patterns in the SITVIT2 database, while 4 shared types containing 8 isolates were newly created. The most prevalent M. tuberculosis lineages were: Haarlem (23/95, 24.2%), ill-defined T superfamily (22/95, 23.2%), the Turkey family (19/95, 20%; previously designated as LAM7-TUR), Beijing (6/95, 6.3%), and Latin-America & Mediterranean (LAM, 5/95 or 5.3%), followed by Manu (3/95, 3.2%) and S (1/95, 1%) lineages. Four of the six Beijing family isolates (66.7%) were MDR. A combination of IS6110-RFLP and spoligotyping reduced the clustering rate from 79% to 11.5% among the drug resistant isolates. CONCLUSIONS: The results obtained showed that ill-defined T, Haarlem, the Turkey family (previously designated as LAM7-TUR family with high phylogeographical specifity for Turkey), Beijing and LAM were predominant lineages observed in almost 80% of the drug-Resistant M. tuberculosis complex clinical isolates in Ankara, Turkey.

[Quality assessment of microscopic examination in tuberculosis diagnostic laboratories: a preliminary study]. / Tüberküloz tani laboratuvarlarinda mikroskopi kalite degerlendirme pilot uygulamasi

Recently, the diagnosis of pulmonary tuberculosis (TB) has based on smear microscopy in the Direct Observed Treatment Strategy (DOTS) programme which provides the basis of treatment worldwide. Microscopic detection of AFB (Acid-Fast Bacilli) is one of the main components in the National TB Control Programmes (NTCP). Precision level in microscopy procedures and evaluations are the most important steps for accurate diagnosis of the disease and to initiate proper treatment. Therefore, the external quality assessment (EQA) is the most important implement to provide the reliability and validity of tests. In countries where NTCP are performed, this task is fulfilled by the National Reference Laboratories (NRL) according to the guidelines of the World Health Organization (WHO). For this purpose a pilot study was initiated by the central NRL of Turkey for EQA of AFB smear microscopy as part of the NTCP on January 1, 2005. A total of 5 laboratories of which 2 were district TB laboratories (A, B), 2 were tuberculosis control dispensaries (C, D), 1 was a national reference laboratory (E), participated in this study. Blind re-checking method (re-examination of randomly selected slides) was used for the evaluation, and the slides were sent to the central NRL with 3 months interval, four times a year, selected according to LQAS (Lot Quality Assurance Sampling) guides. In the re-evaluation of the slides, false positivity (FP), false negativity (FN) and quantification errors (QE) were noted. Laboratory A, sent totally 525 slides between January 1, 2005 and April 1, 2008. In the result of re-checking, 514 (97.9%) slides were found concordant, and 11 (2.1%) were discordant (10 FP, 1 FN). Laboratory B, participated in the study between October 1, 2005 and July 1, 2006 and of the 67 re-examined slides, 60 (89.5%) were concordant and 7 (10.5%) were discordant (2 FP, 0 FN, 5 QE). Laboratory C, sent 235 slides between January 1, 2005 and April 1, 2006; of them 218 (92.8%) were detected as compatible and 17 (7.2%) slides were incompatible (4 FP, 9 FN, 4 QE). Laboratory D, participated in QC for only once between January 1, 2008 and April 1, 2008; and all the 50 slides were found compatible, with no FP, FN and QE. Laboratory E, was included in the study between January 1, 2005 and January 1, 2008 and of the 696 re-checked slides, 690 (99.1%) were reported as compatible and 6 (0.9%) were incompatible (3 FN, 3 QE). Following EQA, on-site evaluation of the laboratories with major errors, was performed and necessary adjustments and training were done. In conclusion, external quality control measures for AFB microscopy is crucial and essential for the tuberculosis laboratory performances for accurate and reliable results.

[Evaluation of Quantiferon-TB Gold and tuberculin skin test in patients with tuberculosis, close contact of patients, health care workers and tuberculosis laboratory personnel]. / Tüberkülozlu hastalar, yakin temaslilari, saglik çalisanlari ve tüberküloz laboratuvari personelinde Quantiferon-TB Gold ve tüberkülin cilt testinin degerlendirilmesi.

Tuberculin skin test (TST) has been used effectively for a long time, despite inherent sensitivity and specificity limitations. Patients with a positive TST without active tuberculosis are identified as having latent tuberculosis infection. Identifying patients with latent tuberculosis infection with this test is an important part of control of the disease. A whole-blood inferferon gamma (IFN-γ) assay, the Quantiferon TB Gold test (QTG; Cellestis, Australia) which is a promising in vitro diagnostic test for the identification of latent tuberculosis infection (LTBI), has potential advantages over the TST. This test includes Myobacterium tuberculosis specific ESAT- 6 and CFP-10 antigens. The aim of this study was to compare the results obtained by QTG and TST in active tuberculosis (TB) patients, close contacts of patients, health care workers and tuberculosis laboratory personel. Twenty-six patients with active pulmonary TB, 6 close contacts of those patients, 11 health care workers with contact to TB patients and 8 TB reference laboratory personnel were included in the study. Prior to administration of the TST, blood samples were drawn from each participant for QTG test. All subjects were asked for BCG vaccination history and examined for a BCG scar. All individuals had a BCG scar. The QTG assay was performed in whole blood samples according to manufacturer's instructions. The agreement between TST and QTG was measured with kappa statistical analysis. In active TB patients (true-infected cases) TST (PPD) positivity was found 34.6% (9/26) while QTG positivity was 65.3% (17/26). Although the positivity rate was higher in QTG test, this difference was not found statistically significant (p > 0.001). TST and QTG positivity rates for health care workers, close house contact of TB patients and TB laboratory staff were as follows, respectively; 36% (4/11) and 27% (3/11); 16.6% (1/6) and 83% (5/6); 37.5% (3/8) and 75% (6/8). The mean PPD diameter was 11 mm in QTG negative group and 14 mm in QTG positive group with a statistically significant difference (p < 0.001). However, there was no statistical significance between QTG positive and negative groups by means of age (p ≥ 0.05) and gender (p < 0.001). In conclusion, QTG assay was superior to TST in its ability to detect LTBI and active TB infection, not to be affected with BCG vaccination, to discriminate responses due to non-tuberculous mycobacteria, and to avoid variability and subjectivity associated with application and reading the TST. Besides, QTG assay needs only one visit to the test unit. However, its being expensive than TST and requirement for special equipments and skilled laboratory personnel, are among the disadvantages of QTG assay.

[Identification and isolation of non-tuberculous mycobacteria from environmental samples]. / Çevre örneklerinden tüberküloz disi mikobakterilerin izolasyonu ve tanimlanmasi

Non-tuberculous mycobacteria (NTM) found frequently in tap water and environment cause important opportunistic infections in immunocompromised patients. The aim of this study was to isolate and identify non-tuberculous mycobacteria in soil, raw milk and water distribution system samples in Mersin (a province located at Mediterranean region of Turkey). A total of 101 water, 124 soil and 40 milk samples collected from the central part and suburban parts of Mersin during November 2003-May 2004 period were included in the study. Water samples were collected from 29 different water distribution systems; soil samples from different parks and gardens and milk samples from raw milks sold at different districts. After the samples were processed by homogenization and decontamination, acid-fast staining and culture into Löwenstein-Jensen medium were performed. Acid-fast bacilli isolated from culture medium were identified by using conventional methods, polymerase chain reaction (PCR)-RFLP (Restriction Fragment Length Polymorphism) and INNO-LIPA Mycobacteria methods. NTM were identified from 4.9% (5/101) of water samples and 0.8% (1/124) of soil samples by culture and PCR. No NTM were detected in the raw milk samples. Three of the NTM strains isolated from water samples were defined as Mycobacterium chelonae type III and two as Mycobacterium kansasii type II. One NTM strain isolated from soil was defined as Mycobacterium fortuitum. It was of note that two of the five NTM positive water samples were tap water samples collected from hospitals. It was concluded that NTM colonization/contamination of water and environment in the hospitals was a potential risk factor in terms of nosocomial infections. Thus surveillance cultures of the water systems and the medical devices in the hospital are necessary to fix the source of NTM, to identify and type the strains and to establish effective control measures such as sterilization, disinfection, maintenance and modernization of water systems.

[Resistance rates to major anti-tuberculosis drugs in Mycobacterium tuberculosis strains isolated from seven different regions of Turkey in 2003-2006 period]. / Ulkemizin yedi farkli bölgesinden 2003-2006 yillari arasinda izole edilen mycobacterium tuberculosis suslarinin majör antitüberkoloz ilaçlara direnç oranlari.

Multi-drug resistance in Mycobacterium tuberculosis (MDR-TB) is a global problem and has increased especially in areas where tuberculosis control programmes are inefficient. The aim of this study was to detect the resistance rates against isoniazide (INH), rifampisin (RMP), ethambutol (EMB) and streptomycin (SM) in M. tuberculosis strains collected from 7 different regions including 62 cities and sent to Refik Saydam Hygiene Center National Tuberculosis Reference and Research Laboratory. Of the patients included, 7.61% were children, 92.39% were adults; 76.16% were male and 23.84% were female. These strains were isolated from sputum (n = 885, 81.11%), gastric lavage (n = 49, 4.49%), pleural fluid (n = 43, 3.94%), urine (n = 30, 2.74%), bronchoalveolar lavage (n = 22, 2.01%), and other clinical samples (n = 62, 8.46%) such as cerebrospinal fluid, lymph node, abscess material, lung tissue. The susceptibilities of the 1091 M. tuberculosis strains against the major anti-tuberculosis drugs were determined by the proportion method in Lowenstein-Jensen medium. Three hundred ninety two of the isolates were from Central Anatolia, 146 from Black Sea, 419 from Aegean, 28 from Mediterranean, 20 from Marmara, 64 from Eastern Anatolia and 21 from South Eastern Anatolia regions of Turkey. The distribution of the strains according to years were as follows: 88 in 2003, 114 in 2004, 341 in 2005 and 548 in 2006. Resistance to at least one of the drugs tested was found in 264 (14.20%) strains. Overall drug resistance rates to INH, RMP, EMB and SM were 12.3%, 10.1%, 6% and 15.8%, respectively. MDR-TB rate was 10% for this 4 years study period. MDR-TB rate was detected as 13.2%, 9.7%, 10.1% and 9.6% in children, adult, male and female patients, respectively. MDR-TB rate did not exhibit a statistically significant difference in terms of sex, age and study years (p > 0.05). However, this rate showed statistically significant difference in terms of geographical regions (p < 0.05). This study emphasized that regular surveillance of M. tuberculosis resistance to the major anti-tuberculosis drugs will provide valuable data for the effective national control and treatment of tuberculosis.

[Susceptibilities of Mycobacterium tuberculosis strains collected from regional tuberculosis laboratories to major antituberculous drugs]. / Bölge tüberküloz laboratuvarlarindan gonderilen Mycobacterium tuberculosis suslarinin majör antitüberküloz ilaçlara duyarliliklari.

The aim of this study was to investigate the susceptibility rates of Mycobacterium tuberculosis strains sent to Refik Saydam Hygiene Center, Tuberculosis Reference and Research Laboratory, Ankara, from seven different regional tuberculosis laboratories between the 1999-2002 period against major antituberculous drugs. The sensitivities of a total 505 M. tuberculosis strains to isoniazid (INAH), rifampicin (RIF), streptomycin (SM) and ethambutol (EMB) were determined by using proportion method in Lowenstein-Jensen medium. Of the strains, 385 (76.2%) were found sensitive to all of the tested drugs, while 120 strains were resistant to at least one of the antituberculous drugs. The resistant strains showed 14 different resistance patterns. The resistance rates were detected as 13.3% for INAH and RIF (67 strains of each), 9.1% for SM (46 strains), and 3.4% (17 strains) for EMB. Multidrug resistant (INAH+RIF) M. tuberculosis was 7.9% (40 strains). The highest resistance rate to INAH, RIF and EMB (21.2%, 21.2% and 10.6%, respectively) was detected in the isolates which were sent from Bursa province (located in northwestern Turkey); the highest SM (18.8%) and multidrug resistance (INAH+RIF) rates (18.8% and 10.6%, respectively) were detected in the strains sent from Elazig and Van provinces (both located in eastern Turkey). Since the inappropriate use of the first and second line antituberculous drugs leads to the development and spread of the resistant strains, "Directly Observed Therapy Shortcourse (DOTS)" is a very important practice. Therefore regional tuberculosis laboratories should be worth considering as the chains of a well-organized national laboratory network, in order to detect the antituberculous drug resistance patterns of the M. tuberculosis strains over the country.

Helicobacter pylori in bronchiectasis: a polymerase chain reaction assay in bronchoalveolar lavage fluid and bronchiectatic lung tissue.

BACKGROUND: A number of studies have implicated an association between H. pylori and diverse extra-gastroduodenal pathologies. Chronic inflammation and increased immune response have been observed in bronchiectasis, likely gastroduodenal inflammatory diseases. H. pylori has been found in the trachea-bronchial aspirates of mechanically ventilated patients. Furthermore, the seroprevalence of H. pylori was found to be significantly higher in patients with bronchiectasis than in the control group. The present study was performed to investigate the possible role of H. pylori in the pathogenesis of bronchiectasis. METHODS: Prospectively, bronchoalveolar lavage fluid (BALF) was obtained from patients with bronchiectasis (n=26) and control (n=20). BALF was subjected to polymerase chain reaction (PCR) to determine the presence of H. pylori and serum IgG against H. pylori was determined with micro-ELISA kit. In addition, PCR was performed to determine H. pylori in surgically removed lung tissues from patients with bronchiectasis (n=97). RESULTS: H. pylori DNA was not detected in the BALF or in lung tissue samples. In addition, anti-H. pylori IgG level in patients with bronchiectasis did not show statistically significant difference from that of the control. CONCLUSIONS: Our study provided evidence that there might be no direct association between H. pylori and bronchiectasis; however, the indirect role of soluble products of H. pylori could not be excluded.

Serum neopterin concentrations in healthy healthcare workers compared with healthy controls and patients with pulmonary tuberculosis.

BACKGROUND: The aim of this study was to compare serum neopterin levels in patients with active pulmonary tuberculosis, healthy healthcare workers who had close contact with patients, and healthy volunteers. All of the healthy volunteers stated that they had not encountered possible risk factors for exposure to Mycobacterium tuberculosis infection. MATERIAL/METHODS: Thirty-nine patients, 39 healthy healthcare workers, and a control group of 39 healthy volunteers who had no infection or other diseases were included in this study. Neopterin assay was performed in accordance with the manufacturer's instructions. RESULTS: Serum neopterin levels were 18.6+/-14.2 nmol/l in patients, 9.8+/-2.9 nmol/l in healthy healthcare workers, and 3.4+/-5.2 nmol/l in healthy volunteers. Serum neopterin levels in each group were significantly different from each other (p<0.01). CONCLUSIONS: Levels of neopterin in patients and healthy healthcare workers significantly differ from the levels in healthy controls. The higher serum neopterin levels in healthy healthcare workers may be attributed to latent Mycobacterium tuberculosis infection, but it does not seem to be used in the diagnosis of latent Mycobacterium tuberculosis infection alone. However, more experiences are needed.

Serum adenosine deaminase enzyme and plasma platelet factor 4 activities in active pulmonary tuberculosis, HIV-seropositive subjects and cancer patients.

The aim of this study was to determine the serum adenosine deaminase (ADA) and plasma platelet factor (PF-4) activities in patients with active pulmonary tuberculosis, HIV seropositive subjects, cancer patients (acute and chronic type lymphoblastic leukaemia) and to compare them with the results of healthy individuals. Eighty-eight subjects were enrolled in this study, 24 patients with active pulmonary tuberculosis, 20 patients with HIV seropositive subjects, 24 patients with cancer, 12 patients with acute type lymphoblastic leukaemia, 12 patients with chronic type lymphoblastic leukaemia) patients and 20 healthy individuals. ADA activity was determined in serum samples using colorimetric method and plasma PF4 activity was measured by using a sandwich-type enzyme immunoassay. When all study groups were compared with the control group, mean serum ADA activities were found to be significantly (p<0.01) higher in the sera of patients with active pulmonary tuberculosis (median, range: 39 IU/l), HIV seropositive subjects (median, range: 31 IU/l) than in the sera of cancer patients (median, range: 15) and healthy controls (median, range: 32 IU/l). Plasma PF-4 activities in active pulmonary tuberculosis patients (median, range: 84 IU/ml) were found to be significantly elevated when compared to HIV seropositive subjects (median, range: 59 IU/ml), cancer patients (median, range 55 IU/ml) and healthy individuals (median, range: 56 IU/ml) (p<0.01). Serum ADA and plasma PF-4 activities showed significant alteration in patients with active pulmonary tuberculosis compared to patients with HIV seropositive subjects, cancer patients and healthy individuals. In conclusion, we suggest that serum ADA and PF-4 activities can be used in the diagnosis of tuberculosis as an supplementary laboratory test in combination with clinical and laboratory findings. Further controlled studies are necessary to determine the importance of the PF-4 and ADA activities in patients with active pulmonary tuberculosis, HIV seropositive subjects and cancer patients.

[Retrospective evaluation of Cobas Amplicor system in the rapid diagnosis of Mycobacterium tuberculosis complex]. / Mycobacterium tuberculosis kompleksinin hizli tanisinda cobas amplicor sisteminin retrospektif olarak degerlendirilmesi.

The aim of this study was to evaluate the value of Cobas Amplicor Mycobacterium tuberculosis (MTB) polymerase chain reaction (PCR) system in the rapid diagnosis of tuberculosis. During the study period, the results of acid-fast stain (AFS), culture and Cobas Amplicor MTB system obtained from 937 clinical (158 respiratory and 779 non-respiratory) specimens were retrospectively evaluated. When culture results were accepted as the gold standard, the sensitivity, specificity, positive and negative predictive values of Cobas Amplicor MTB PCR system were found as 100% for smear-positive respiratory specimens, 75%, 91.7%, 40% and 98% for smear-negative respiratory specimens, and 89.5%, 91.8%, 65.4% and 98.1% for all of the respiratory specimens, respectively. These rates were found as follows for smear-positive, smear-negative and all of the non-respiratory specimens, respectively; 100%, 66.7%, 83.3% and 100%; 29.2%, 97.3%, 26.9% and 97.6%; and 50%, 97.1%, 44.7% and 97.6%. The overall inhibition rate for Cobas Amplicor MTB was 4.8%. In conclusion, Cobas Amplicor MTB PCR system was considered as a rapid and reliable method for the diagnosis of tuberculosis in smear positive pulmonary samples. However, in smear negative pulmonary samples and non-respiratory samples, test results should be evaluated together with clinical and other laboratory data.

[Evaluation of a commercial enzyme immunoassay for the detection of interferon gamma levels in active tuberculosis patients and vaccinated healthy subjects]. / Aktif tüoberkülozlu hastalar ve asili saglikli bireylerde interferon gamma düzeylerinin saptanmasinda ticari bir enzim immon yonteminin degerlendirilmesi.

The detection of plasma interferon gamma (IFN-g) levels has an important value for the evaluation of cell mediated immune response to Mycobacterium tuberculosis. The aim of this study was to investigate the plasma IFN-g levels by a commercial enzyme immunoassay (ELISA) and to compare the levels between recently diagnosed culture positive lung tuberculosis patients and BCG vaccinated healthy controls. Twenty-three patients with active lung tuberculosis (13 males, 10 females) and 34 BCG vaccinated healthy adults (16 male, 18 female) have been included in the study. The control subjects were questioned about passed tuberculosis infection and/or a contact with tuberculosis patients. No risk factors for exposure to M. tuberculosis were found in the control group. IFN-g levels were measured by QuantiFERON-TB (Cellestis, Australia) kit, and 22 of patients and 17 of control subjects were found to be positive. As a result, the sensitivity of QuantiFERON-TB test was high (95.6%), however its specificity was quite low (50%). In conclusion, QuantiFERON-TB may be used as a supplementary diagnostic test in patients considered to have active tuberculosis, before treatment. As BCG is in routine vaccination programme and the number of active tuberculosis cases is high in our country, this test seems to be invalid for the diagnosis of latent tuberculosis. Therefore, more specific tests that are not affected by the vaccine response, are required for the diagnosis of latent tuberculosis.

[Microbiological evaluation of the cases prediagnosed as pulmonary tuberculosis based on clinical and radiological findings]. / Klinik ve radyolojik bulgular ile akciger tüberkülozu ön tanisi alan hastalarin mikrobiyolojik sonuçlari.

In the present study, 34 new pulmonary tuberculosis cases prediagnosed upon clinical and radiological findings, have been evaluated by means of microbiological aspects such as microscopy [Ehrlich-Ziehl Neelsen (EZN) and Auramin-Rhodamine (A-R) stains], culture [inoculation into Lowenstein-Jensen (LJ) media] and three different molecular methods [Cobas Amplicor Mycobacterium tuberculosis polymerase chain reaction (PCR), Gen-Probe amplified M. tuberculosis direct test and TaqMan Real Time ABI 5700 PCR]. M.tuberculosis positivity of the sputum samples were 55.8% (n: 19) with EZN staining, 52.9% (n: 18) with A-R staining and 58.8% (n: 20) with culture methods. Among molecular techniques, Cobas Amplicor and TaqMan PCR yielded positive results in 21 patients (61.7%), however, the positivity rate of Gen-Probe AMTD was 47.0% (n: 16). Of 34 samples, 10 (29.4%) were found to be positive with all of the methods used, while six (17.6%) of them were negative with all methods. It can be concluded that molecular methods may aid to the diagnosis of tuberculosis, in conjunction with clinical findings and bacteriological results.

[Comparison of auramine-rhodamine and Erlich-Ziehl-Neelsen staining methods for the diagnosis of tuberculosis]. / Tüberküloz tanisinda auramine-rhodamine ve Erlich-Ziehl-Neelsen boyama yöntemlerinin karsilastirilmasi.

The aim of this study was to compare auramin-rhodamine (A-R) and Erlich-Ziehl-Neelsen (EZN) staining techniques for the diagnosis of tuberculosis. Of 311 sputum samples collected from active pulmonary tuberculosis patients and tuberculosis-suspected patients, 103 (33%) were found culture positive. In the direct microscopic examination of EZN stained smears, 86.4% (89/103) of culture positive samples, and 3.8% (8/208) of culture negative samples yielded asido-resistant bacteria, while these rates were 74.8% (77/103) and 11.5% (24/208) for A-R staining method. When culture was accepted as reference method, the specificity and sensitivity of the staining techniques were found as 88.5% and 74.8% for A-R, and 96.2% and 86.4% for EZN, respectively. As a result, it was concluded that, the use of A-R staining alone, could not be an alternative method to EZN staining.