423. What is new in the diagnosis of tuberculosis? T , S
Transcription
423. What is new in the diagnosis of tuberculosis? T , S
Thematic Poster Session Hall 3-53 - 12:50-14:40 T UESDAY, S EPTEMBER 21 ST 2010 423. What is new in the diagnosis of tuberculosis? P4987 Diagnosis of smear negative pulmonary tuberculosis (PTB) by pooling two induced sputums on the same day Mei Ying Chew, Jason Phua, Adeline Tan, Vimal Palanichamy, T.K Lim. Medicine, National University Health System, Singapore, Singapore Background: The most cost effective method to diagnose smear negative PTB is repeat testing of inducted sputum over several days. Aim: The aim of this study was to evaluate the diagnostic yield of two induced sputum specimens pooled together over 1 day for microbiologic testing in patients with smear negative PTB. This may increase the speed of diagnosis and decrease the workload of laboratories. Methods: We enrolled consecutive adults hospitalized with suspected PTB who were unable to expectorate or whose sputum was acid fast smear negative. Two sputum inductions were obtained on the first day at least 4 hours apart and one more induction was performed on the second day. The 2 specimens on the first day were pooled together and tested for acid fast smear and cultures. The third specimen on the second day was tested separately. The final diagnosis of PTB was based on clinician diagnosis and a positive culture from all available tests. Results: Among the first 50 patients, 150 sputum inductions were performed without any adverse events. 15 out of 50 patients (30%) had confirmed final 922s Abstract printing supported by Chiesi Farmaceutici SpA. Visit Chiesi Farmaceutici SpA. at Stand G.90 Thematic Poster Session Hall 3-53 - 12:50-14:40 T UESDAY, S EPTEMBER 21 ST 2010 diagnosis of PTB. There were 12/15 positive smears and 15/15 positive cultures in the pooled specimens. There were 9/15 positive smears and 13/15 positive cultures in the third specimens. These specimens did not detect any additional cases. Conclusions: Microbiological testing of two pooled induced sputum examinations performed on the same day is practical and effective in diagnosing smear negative PTB. There may be little additional yield from further repeating the sputum induction on subsequent days. P4988 New immunological skin test in differential diagnostics of tuberculosis and nonspecific diseases of lungs Tatiana Morozova 1 , Tatiana Salina 2 . 1 Phthisiology, Medical University, Saratov, Russian Federation; 2 Phthisiology, Medical University, Saratov, Russian Federation The aim of this research was increase efficiency diagnostics of tuberculosis. We checked 73 patients at the age from 17 till 80 years with difficulties of verification of the diagnosis of tuberculosis and 35 healthy persons, who have had professional contacts with TB infection, but without local changes in bodies. We studied the effectiveness of the new immunological method Diaskin-test (DST), based on intradermal injection of the drug, which contains two specific antigens ESAT-6 and CFP-10 (“Lecco”, Russia) at a dose of 0.2 mg in 0.1 ml. We evaluated result reaction visually after 72 hours and measured the size of induration in mm. In patients with active pulmonary TB is received positive result DST in 47 cases and the sensitivity of a method has made 92,2%. The average size of induration in mm in these patients was 16,4±1,0 mm, median – 17, moda – 18, which was significantly greater than in patients with nonspecific lung diseases – 3,6±1,2 mm, median – 2, moda – 0 (p1-2=0,00014) and healthy – 3,97±1,1 mm, a median – 2, a moda – 0 (p1-3=0,00012). In the group of patients with nontuberculous lung diseases (sarcoidosis, innate and parasitic cysts, lung cancer, etc.) received 4 false-positive result and specificity of the method in this group was 81,9%, and who have had professional contacts with TB infection – 4 (88,6%). Diagnostic efficiency of the method – 87,1%. Thus, the new Diaskin-test is highly sensitive and highly specific method for detection of active TB infection. P4989 An immunochromatographic rapid test for Mycobacterium identification Elena Cojocaru 1 , Beatrice Irina Saramet 2 , Daniela Diculencu 2 . 1 University of Medicine and Pharmacy, Faculty of Medicine, Department of Physiology, Iasi, Romania; 2 Clinic of Pulmonary Diseases, Microbiology Laboratory, Iasi, Romania In Romania, the M. tuberculosis identification is based on the phenotipical features (niacin, reductase, catalase reactions). Thus, a new Immunochromatographic Rapid Test TB Ag MPT 64 seems to be suitable for a rapid differentiation between tuberculosis and non-tuberculosis mycobacteria. Aim: Due to the lack of the modern identification tests for M. tuberculosis the Immunochromatographic Rapid Test TB Ag MPT 64 could be useful for rapid and cost effective mycobacteria identification. We intended to evaluate if this test is suitable for the mycobacterial identifications. Methods: The Immunochromatographic Rapid Test for M. tuberculosis complex identification use mouse monoclonal anti-MPT 64. This test kit can be easily used for rapid identification of the M. tuberculosis complex in combination with culture systems based on liquid media. The test is available on the market for professional use. All cases were selected from the Clinic of Pulmonary Disease, Iasi, Romania. The tests were performed according to the producer’s recommendation. Results: We have tested 25 cultures identified in the Microbiology Laboratory between November and December 2009. The Immunochromatographic Rapid Test was positive in 52% of cultures tested. All positive tests were identified M. tuberculosis based on the phenotipical features. From negative tests, using laboratory techniques 41.7% were M. non-tuberculosis, since 58.3% were classified as M. tuberculosis. Conclusion: In this population study, this Immunochromatographic Rapid Test based on Ag MPT 64 seems to be suitable for culture identification. Further trials are needed to confirm this early findings by using new methods which are not still available in Romania. Objectives and methods: Using enzyme-linked immunoassay developed by Mona Ltd. (Moscow) for measuring IFN-γ cell response to MTB antigens (tuberculin PPD and specific antigen ESAT-6) in whole blood samples after 24 hour incubation, we have studied 2 groups: 60 TB patients aged 19-66 yrs. and 67 patients with morphologically identified sarcoidosis aged 19-60 yrs. Results: In patients with sarcoidosis IFN-γ levels in whole blood was significantly lower than in patients with TB: the respective 95% CIs for the response to PPD were 33,2-100,2 pg/mL versus 236,9-367,4 pg/mL (P <0,001 Mann-Whitney U-test); while the respective 95% CIs to ESAT-6 were 3,3-15,2 pg/mL versus 59,3-126,3 pg/mL (P< 0,0005 Mann-Whitney U-test). When using 70 pg/mL as a cut-off point, 97% patients with sarcoidosis had no IFN-γ response to ESAT-6 and 76,1% patients with sarcoidosis had no IFN-γ response to PPD as compared with 80% TB patients who had such a response. The diagnostic efficacy of this test is 77,9%. In case of IFN-γ release lower than 70 pg/mL odds ratio for sarcoidosis is 12,8. The positive predictive value is 81% [95% CI 72,1/87,9] and the negative predictive value is 75% [95% CI 66,3/81,8%], p<0,001. Conclusion: The antigen-specific IFN-γ release assay may be used for the differential diagnosis between TB and sarcoidosis. P4991 Can plain chest radiography and age accurately predict the presence of smear positive pulmonary tuberculosis? Rifat Rashid, Amanda James, Heinke Kunst. Respiratory Medicine, Heartlands Hospital, Birmingham, West Midlands, United Kingdom Rationale: Delayed diagnosis of smear-positive pulmonary tuberculosis (PTB) is common and believed to contribute significantly to transmission. Persons with symptoms suggestive of PTB often have CXRs prior to sputum examination and clinic assessment. It is unclear if chest radiography and age are markers of smear positive PTB. Methods: We conducted a prospective study including consecutive patients with features suggestive of active PTB on chest radiograph referred by radiologists to our institution from November 2008 to November 2009. Patients with typical changes of PTB on CXR were compared with those with atypical changes. Typical changes include upper zone consolidation and cavitation. Logistic regression analysis was performed to assess whether age (< 30 years vs >30 years) was an independent predictor of sputum smear positivity. Results: 54 patients had CXR changes consistent with focal or lobar consolidation, 20 patients were found to be smear positive. Younger patients were more likely to be smear positive (p= 0.028, odds ratio 0.4). In addition patients with typical changes of PTB were more likely to be smear-positive (p=0.17, OR 1.36). Conclusion: Young patients with focal or lobar upper zone changes on chest radiography suggestive of PTB should be referred to a TB clinic to exclude the presence of smear-positive pulmonary TB as a matter of urgency. P4992 Interferon-gamma responses and the spectrum of tuberculosis Graham Bothamley, William Ricketts. Respiratory Medicine, Homerton University Hospital, London, United Kingdom Introduction: Th1 responses are considered more important in primary tuberculosis and Th2 in post-primary, pulmonary disease (PTB). Vitamin D may be important in cellular immunity. We wished to know whether interferon-gamma levels varied with the site of tuberculosis and/or vitamin D levels. Method: Patients with tuberculosis were offered an interferon-gamma release assay (IGRA: QuantiFERON Gold in tube) and vitamin D levels were measured. The diagnosis of PTB was by culture and of other forms of disease by culture, histology or clinical with a positive tuberculin skin test and IGRA. Student’s t-test was used to compare interferon-gamma levels. Results: 48 patients were entered (all HIV-negative). All PTB were culture-positive and 13 were also smear-positive. Other primary TB consisted of 3 psoas abscesses, 1 spinal, 1 rib, 6 with disseminated disease and 2 with abdominal TB. Those with mediastinal lymph node TB (MLN) had the highest levels (p=0.01 compared with extrathoracic lymphadenitis (LN) or PTB). Other primary disease had the lowest levels (p=0.03 compared to PTB). Test P4990 The significance of the antigen-specific IFN-γ release assay in vitro for differential diagnosis between sarcoidosis and tuberculosis (TB) patients Natalya Makarova 1 , Sergey Borisov 3 , Tatyana Vlasik 2 , Elena Yanushevskaya 2 , Natalya Kashirina 2 , Evgeny Efremov 2 , Galina Ignashenkova 2 , Mikhail Vladimirskij 1 . 1 Diagnostic, Phthisiopulmonology Research Institute I.M. Sechenovs Moscow Medical Academy, Russian Federation; 2 Laboratory, Mona Ltd., Moscow, Russian Federation; 3 Health Department, Scientific and Clinical TB Center of Moscow Government, Russian Federation IGRA (Log IU/mL) Vit D (n > 50 nmol/L) MLN n=5 LN n=12 PTB n=18 Other primary n=13 1.06±0.10 0 0.70±0.12 1 0.68±0.09 2 0.48±0.18 0 5 other patients with MLN had IGRA titres of 9.2, 12.6, 12.0, 12.7 and 11 IU/mL. There was no correlation of IGRA titres with vitamin D levels. The one LN patient with normal vitamin D had a titre of 9.4 IU/mL (2.3 SE above the log mean), while in PTB the two titres were around the mean (6.6 & 4.2 IU/mL). Conclusion: IGRA responses are highest in those with MLN and lowest in those with other primary disease. Background: The interferon-gamma (IFN-γ) is known to be essential for a protective immunity to tuberculosis (TB) and plays an important role in a granulomas formation. The role of M.tuberculosis (MTB) antigens in the sarcoidosis pathogenesis is still unclear, while the granulomatous reaction is similar with that in TB. 923s Abstract printing supported by Chiesi Farmaceutici SpA. Visit Chiesi Farmaceutici SpA. at Stand G.90 Thematic Poster Session Hall 3-53 - 12:50-14:40 T UESDAY, S EPTEMBER 21 ST 2010 P4993 Efficacy of post bronchoscopic sputum smear in diagnosis of tuberculosis, a survey Majid Malekmohammad, Majid Marjani, Parvaneh Baghaei, Zahra Sadr. Pulomonology, Masih Daneshvary Hospital, Tehran, Islamic Republic of Iran Reactivity of multiantigen cocktails was greater then that of any single antigen, even when the same amount of total protein was used for cocktails and for each single antigen (total antigen area response compared with that of the reference was 65-81.3% for singles and 124% for cocktail). The results open the way to formulate a new specific skin test for accurate diagnosis and TB decrease. Introduction: Diagnosis of tuberculosis may become a major problem for both the patient and management team. Aim: To identify the sensitivity of post-bronchoscopy sputum smear in patients with a suspicious diagnosis of tuberculosis. Methods: In a prospective study, we analyzed 126 patients suspicious of having tuberculosis without bacteriologic confirmation. Sputum smears, bronchoscopy with bronchial washings, and post bronchoscopy sputum smears were performed, respectively. Results: 126 patients were studied between December 2008 and December 2009. 32 patients (25.4%) with negative initial sputum smears had positive bronchial washings for acid fast bacilli. Also, in 15 (58%) patients diagnosis of tuberculosis was based on post bronchoscopy sputum analysis.Moreover, 94 (27.7%) patients with a final diagnosis of tuberculosis had negative post-bronchosopic sputum smears. On the other hand, among 32 patients with positive bronchial washings, 28 (87.5%) had positive sputum smears after fiberoptic bronchoscopy. Conclusions: Sensitivity of post-bronchoscopy sputum analysis for tuberculosis is 58%. Classification of these patients as smear negative tuberculosis is doubtful. P4994 Serum adenosine deaminase (ADA) activities and neopterine levels during therapy in patients with pulmonary tuberculosis and community-acquired pneumonia Remise Gelisgen 1 , Hafize Uzun 1 , Füsun Erdenen 2 , Ozlem Kar 2 , Omur Tabak 2 , Gungor Camsari 3 , Bilge Ozdemir 2 . 1 Biochemistry, Istanbul University, Cerrahpasa Medical Faculty, Turkey; 2 Internal Medicine, Istanbul Education and Research Hospital, Turkey; 3 Yedikule Chest Disease and Chest Surgery, Education and Research Hospital, Istanbul, Turkey We investigated the effect of therapy on serum ADA activities and neopterine levels in patients with pulmonary tuberculosis and community-acquired pneumonia. 17 tuberculosis, 24 pneumonia, and a control group of 20 healthy volunteers were included in this study. ADA and neopterin assay were performed by colorimetrically and ELISA, respectively, before and two months after therapy (isoniazid, rifampin, ethambutol, and pyrazinamide) of patients with tuberculosis and before and 15 days after therapy (ampicilline-sulbactam) of pneumonia. Serum ADA activities and neopterin levels were significantly higher in patients with tuberculosis than those in pneumonia and controls. Serum neopterin levels were significantly higher in patients with pneumonia than controls. No significant differences were found in ADA activities between pneumonia and control subjects. Serum ADA activities and neopterin levels significantly decreased in tuberculosis at 2nd month of treatment. Serum neopterin levels significantly decreased in patients with pneumonia after treatment, but ADA activities were not changed. Serum ADA activities and neopterin levels were still significantly higher in patients with tuberculosis than controls despite the treatment. Serum ADA activities and neopterin levels significantly increased in the presence of mycobacterium tuberculosis; combined treatment decreased the serum ADA activities and neopterin levels. Measurements of serum ADA activities and neopterin levels are useful in monitoring drug treatment and immune response to tuberculosis; neopterin also seem to be valuable for pneumonia, whereas ADA activities are not. P4995 Recombinant PPD proteins to be used as specific skin test for tuberculosis Henriette Stavri 1 , Nadia Bucurenci 2 , Karen Dobos 3 , Irina Ulea 1 , Adriana Costache 2 , Agatha Wieczorek 3 . 1 Mycobacterial Antigens Dept., Cantacuzino Institute, Bucharest, Romania; 2 Enzymology and Applied Microbiology, Cantacuzino Institute, Bucharest, Romania; 3 Dept of Microbiology, Immunology,& Pathology, Colorado State University, Ft Collins, Colorado, United States The tuberculin skin test (TST) with PPD IC-65 is used in tuberculosis (TB) diagnosis in Romania. Our aim was to develop a M. tuberculosis (M.tb.) specific skin-test reagent, without false positive results due to BCG vaccination, using recombinant antigens. In order to identify PPD IC-65 proteins, digestion of the whole sample in solution and of gel-excised bands were performed. The proteins have been further analyzed by tandem mass spectrometry and compared with those from PPD S2 (US reference tuberculin standard). We found 11 common proteins. Three of them known as specific antigens for M.tb: MPT64/Rv1980c, ESAT 6/Rv3875 and CFP 10/Rv3874, were cloned and overexpressed in E. coli strains as NH2 terminally polyhistidine tagged fusion proteins and purified as recombinant proteins. To formulate optimal and specific immunodiagnostic cocktails, the recombinant antigens (singles or in mixture) were evaluated by skin testing several guinea pigs groups sensitized with M.tb H37Rv and BCG, as controls. For single antigens as well for the mixture, the best results were obtained using 3 μg/0.1 ml of each recombinant protein, equivalent to 105 TU/0.1ml. Each animal was simultaneously tested with PPD IC-65, 2 TU/0.1 ml, as reference.The controls did not react with any of these antigens but elicited skin test responses with the reference. P4996 Widal test positivity and quinolone use in cases of tuberculosis Tushar Sahasrabudhe, Iqbal Siddiqui, Sandeep Garg. Department of Pulmonary Medicine, Padmashree Dr. D.Y. Patil Medical College, Pune, Mahrashtra, India Both TB and Enteric fever are common in India. TB is the commonest cause of PUO in the country. While evaluating fever, it is a standard practice to order Widal test and if positive, a course of quinolone is given for a period of 2 weeks as a monotherapy. Quinolone resistance in Mycobacteria may develop on short exposure to quinolone monotherapy. We evaluated Widal test positivity and past quinolone use based on the Widal test, in cases of tuberculosis admitted to the pulmonary medicine ward. A control group of patients admitted with diagnosis other than TB and enteric fever was also evaluated to determine if Widal test positivity is significantly different in TB. A total of 31 TB and 29 non-TB patients were evaluated. The pre-hospitalization record of the investigations and treatment for current illness was reviewed. Widal test using tube-test quantitative method was done. Titer of 1:80 with either O or H antigen was taken as a positive test. The Widal test was found positive in 7/31 (22.58%) of TB cases and 9/24 (37.5%) in non-TB cases. This indicates very high false positivity of Widal test in both groups. 7 of 31 (22.58%) of TB patients had already undergone the test before hospitalization and had received a course of quinolone as a monotherapy based on a positive test. Interestingly only 2 of them were found positive during the study. The study points to a situation where the clinician feels justified in using a quinolone monotherapy in undiagnosed cases of TB. This may increase risk of XDR TB. P4997 Comparison of Quanti-FERON-TB Gold and tuberculin skin test for diagnosis of latent tuberculosis infection in patients with chronic schizophrenia Esin Aktas Cetin 1 , Faruk Ciftci 2 , Sinan Guloksuz 3 , Kaya Koksalan 4 , Ogun Sezer 5 , Esat Timuçin Oral 3 , Turan Cetin 3 , Erhan Kurt 3 , Hatice Kaya 6 , Gunnur Deniz 1 . 1 Department of Immunology, Istanbul University, The Institute of Experimental Medicine (DETAE), Turkey; 2 Department of Pulmonary Medicine and Tuberculosis, Gulhane Military Academy of Medicine, Istanbul, Turkey; 3 Department of Psychiatry, Bakirkoy Prof. Mazhar Osman Teaching and Research Hospital, for Psychiatry, Neurology & Neurosurgery, Istanbul, Turkey; 4 Molecular Tuberculosis Epidemiology Laboratory, Istanbul University, The Institute of Experimental Medicine (DETAE), Turkey; 5 Department of Pulmonary Medicine, Microbiology Laboratory, Gulhane Military Academy of Medicine, Istanbul, Turkey; 6 Department of Pulmonary Medicine, Gulhane Military Academy of Medicine, Ankara, Turkey Objective: In this study we aimed to determine the prevalance of latent tuberculosis infection (LTBI) in patients who were hospitalized with diagnosis of chronic schizophrenia and compared the diagnostic value of tuberculin skin test (TST) and Quanti-FERON-TB Gold (QFT-G). Material and method: In our study, M. tuberculosis specific ESAT-6 and CFP-10 antigen specific interferon gamma (IFN-γ) cytokine levels were investigated by QFT-G method (cut off value ≥ 0,35 IU/ml) in male (n=71, mean age 55,12±10,03) and female (n=95, mean age 58,21±11,06) patients who had been hospitalized for at least three years with diagnosis of chronic schizophrenia. All of the patients were on antipsychotic and free of immune-suppresive treatment. TST was performed using 5 TU PPD-RT 23 (TW80) according to Mantoux method and diameter of enduration was measured after 72 hours of inoculation. The patients were evaluated clinically and radiologically. Results: TST was positive in 99 (59,63%) and QFT-G was positive in 88 (53,01%) patients and there was a low agreement between the two tests (κ=0,38). Four patients with active disease were diagnosed baised on radiological and clinical findings and the institution was notified. In this study, the frequencies of both of the active disease (2,4%) and LTBI (53%) were found to be very high. Conclusion: A very high prevalance rate of active tuberculosis and relatively high LTBI rate were observed in our schizophrenic patient group and QFT-G is considered to be a useful diagnostic tool. P4998 Importance of QuantiFERON-TB Gold and tuberculin skin test in the differential diagnosis of active and inactive pulmonary tuberculosis Ekrem Cengiz Seyhan 1 , Kaya Köksalan 2 , Gülsah Günlüoglu 1 , Mustafa Düger 1 , Sinem Nedime Sökücü 1 , Atayla Gençoglu 1 , Erdogan Çetinkaya 1 , Sedat Altin 1 . 1 Chest Disease, Yedikule Chest Disease and Thoracic Surgery Research and Education Hospital, Istanbul, Turkey; 2 Experimental Research Institude, Istanbul University, Turkey Aim: This study is aimed to examine the importance of QuantiFERON-TB Gold test and tuberculin skin test which are based on examination of IFN-γ in the differential diagnosis of active and inactive pulmonary tuberculosis. 924s Abstract printing supported by Chiesi Farmaceutici SpA. Visit Chiesi Farmaceutici SpA. at Stand G.90 Thematic Poster Session Hall 3-53 - 12:50-14:40 T UESDAY, S EPTEMBER 21 ST 2010 Method: 51 Patients with active pulmonary TB and 47 inactive cases were included to the study. After the blood samples were taken for the QuantiFERON-TB Gold test, TST was done and above 15 mm is accepted as positive. Results: QuantiFERON-TB Gold test positivity was determined at 48 of 51 (98%) patients with active TB and at 40 of 47 (85%) patients with inactive TB. There was no difference statistically in terms of QuantiFERON-TB Gold test positivity between two groups. There was no meaningful difference is determined between the patients with active pulmonary TB [4.28 (0.9- 12.9) IU/mL] and the patients with inactive pulmonary TB [3.15 (0.9- 5) IU/mL] in terms of QuantiFERON-TB Gold Nil levels. Meaningful correlation was found between the Nil levels of the cases with inactive TB and the year in which the disease occurred. There was no meaningful difference between the groups in terms of PPD induration diameter and positivity. At the division of active and inactive pulmonary TB the sensitivity was determined as 45% and specificity was determined as 82%, when 3,4 IU/mL was taken as a threshold. Conclusion: TST and QuantiFERON-TB Gold are not determined as valuable in the differential diagnosis of active and inactive pulmonary tuberculosis P4999 Comparison of Lowenstein-Jensen and Bactec-MGIT 960 culture systems in the diagnosis of pulmonary tuberculosis Esra Kadioglu 1 , Leyla Saglam 1 , Esin Aktas 2 , Metin Akgün 1 , Esin Sonkaya 1 , Metin Gorguner 1 . 1 Chest Diseases Department, Ataturk University Medical Faculty, Erzurum, Turkey; 2 Microbiology Department, Ataturk University Medical Faculty, Erzurum, Turkey Aim: We aimed to compare isolation rates and growth time of Mycobacterim Tuberculosis by BACTEC – Mycobacterium Growth Indicator Tube (MGIT) with Lowenstein – Jensen (LJ) culture methods in patients with tuberculosis (TB) suspicion. Method: Asid fast bacilli (AFB) was examined with in 105 sputum samples obtained from the patients suspected TB (% 59 male and % 49 female) stained with Ehrlich-Ziehl-Neelsen (EZN) via direct microscopic detection. LJ and BACTEC – MGIT culture systems were done. SPSS 11 programme was used for stasistically evaluation. Results: Direct smear or culture positivity was determined in 87% of the cases. Of the 91 specimens, fifty cases were male and forty-one female. Average age was 41±2 years old. Positivity rates were 77%, 69%, 92% for AFB smears, LJ and BACTEC – MGIT, respectively. Average detection days of M. Tuberculosis were 39±10 days and 12±6 days for LJ and BACTEC – MGIT, respectively. Growth rates were 66% when combined LJ medium and BACTEC – MGIT systems, 96% either LJ or BACTEC – MGIT and 30% only one of the culture systems. Determination of positivity was more rapidly in BACTEC –MGIT 960 system than LJ (p<0.0001). Conclusion: BACTEC – MGIT 960 system provided rapidly (p<0,0001) and sensitive results for diagnosis of pulmonary TB. Combination of both BACTEC – MGIT and LJ media increased diagnosis rates. P5000 Diagnosing negative sputum tuberculosis using automated MGIT Lina Kayalli 1 , Omar Balash 1 , Abdalla Khoury 2 , Diana Zammar 2 . 1 Department of Bacteriology, Aleppo University Hospital, Aleppo, Syrian Arab Republic; 2 Department of Internal Medicine, Aleppo Faculty of Medicine, Aleppo, Syrian Arab Republic WHO estimates that eight million people get TB every year, of whom 95% live in developing countries. An estimated 3 million people die from TB every year. In developing countries, the impact of HIV infection on the TB situation, especially in the 20-35 age group, is worthy of concern. A final factor contributing to the resurgence of TB is the emergence of multi-drug resistance. Objective: The results of culture of different TB pathological samples using BACTEC MGIT 960 system were subtyped, evaluated and compared with Lowenstein-Jensen (LJ) medium to estimate the recovery rate and time of detection of pathologic mycobacteria. Materials & Methods: This prospective study was done in a tertiary university hospital. A total of 180 negative sputum samples were examined for the presence of mycobacteria. We made ZN stain to all specimens, which were also processed and inoculated separately in the BACTEC MGIT 960 system, and LJ medium. Then we made drug sensitivity test to all positive sputum samples for the first–line drugs (IZN.RIF,PYR,STM, ETH). Results: The BACTEC MGIT 960 detected 135 isolates (75%), followed by LJ with 108 isolates (60%). The mean time of detection was 9.6 days with MGIT 960, 25.1 days with LJ. The contamination rates were 4% with MGIT 960, 7% with LJ. We did the susceptibility testing for Mycobacterium bacilli, with a susceptibility rate for INH 89%, RIF 82%, PYR 94%, ETH 92% and STM 84% respectively. The mean time to MGIT susceptibility testing results was 6.6±0.7 days. Conclusion: We found that the MGIT culture will ameliorate the diagnosis of negative sputum tuberculosis with better control and treat our TB patients. Keywords: ZN: Zeil-Nelson, MGIT: Mycobacteria Growth Indicator Tube P5001 The compare the results of the biochip-assay and BACTEC MGIT 960 Jainagul Isakova 1 , Nurmira Sovhozova 1 , Almaz Aldashev 1 , Zoy Goncharova 2 . 1 Molecular and Cell Biology, Institute of Molecular Biology and Medicine, Bishkek, Kyrgyzstan; 2 Microbiology, National Center of Phthisiology, Bishkek, Kyrgyzstan Multidrug-resistant tuberculosis (MDR-TB) is a serious problem in Kyrgyz Republic. The role of rapid diagnostics of drug resistant strains of M. tuberculosis (M. tb) is critical for the management of patients with MDR-TB. Previously we have shown that the biochip-assay is comparable in sensitivity and specificity to the conventional bacteriological culture methods. Aim: To evaluate the sensitivity and specificity of biochip-assay compared to automated MGIT 960 method. Materials and methods: We analyzed 133 sputum samples from Tb patients using the biochip-assay and BACTEC MGIT 960 approach. The resistance to rifampicin (RIF), isoniazid (INH) and MDR obtained with the BACTEC MGIT 960 system served as the “standard” for comparison to the results obtained with TB-biochip assay. Results: In the case of RIF-resistant samples the sensitivity and specificity were 88% and 96.%, respectively. In the case of IHN resistance, the sensitivity was 93% and specificity also was 93%. For MDR resistant strains, the sensitivity and specificity were 90% and 97%, respectively. Conclusion: Biochip-method has been demonstrated sensitivity and specificity comparable to MGIT 960 approach. At the same time it has the advantage of direct identification of mutations in rpoB, katG, inhA and aphC genes associated with multi drug resistance of. M.Tb. Also biochip-assay is more rapid (2 days) compared to MGIT 960 approach (3 weeks). P5002 Genotyping of M. tuberculosis circulating in penitentiary system of Kyrgyz Republic Jainagul Isakova 1 , Igor Mokrousov 2 , Violetta Valcheva 3 , Nurmira Sovhozova 1 , Almaz Aldashev 1 , Nalin Rastogi 3 . 1 Molecular and Cell Biology, Institute of Molecular Biology and Medicine, Bishkek, Kyrgyzstan; 2 Microbiology, St. Petersburg Pasteur Institute, Russian Federation; 3 TB and Mycobacteria Unit, Institut Pasteur, Guadeloupe, France Multidrug resistant tuberculosis is a major problem in penitentiary system of Kyrgyz Republic. Multidrug resistance (MDR) depends of genotyping of M. Tuberculosis (M.tb). Aim: To determine the contribution of the M. tb Beijing genotype and association with multidrug resistance of M. tb in the penitentiary system of Kyrgyz Republic. Material and methods: DNA samples of M. tb were collected from 56 adult newly diagnosed TB patients in the penitentiary system of Kyrgyzstan. Drug resistance of M. tb to rifampicin (RIF) and isoniazid (INH) were analysed by TB-Biochip assay. IS6110-inverse PCR method, spoligotyping and VNTR (12 MIRU and 3 hypervariable loci) were used for genotyping of M. tb. Results: The Beijing strains of M. tb was detected in 75% (42 of 56), Haarlem – 10,7% (6 of 56) and LAM profiles were detected in 10,7% (6 of 56) of samples. Two samples had a mixture of the Beijing and non-Beijing profiles. We have identified rpoB mutations in 16 (28.5%) of 56 samples. The most prevalent mutation was Ser531→Leu which was identified in 13 of 16 rpoB mutant samples. Three other samples had mutations Leu511→Pro, Asp516→Tyr or His526→Asp and one strain had double mutation Ser531→Leu and Leu511→Pro. Mutations in INH resistance genes were detected in 31 (55.4%) samples almost all of them were in katG gene Ser315→Thr and only one sample had the mutation in the inhA gene promoter region. 15 samples (26.8%) had mutations in both katG/inhA and rpoB genes and were multidrug-resistant strains. In MDR strains 13 of 15 are of Beijing genotype. Conclusion: The Beijing genotype of M.tb have prevailed in penitentiary system of Kyrgyzstan and associated with MDR. P5003 Comparison of tuberculin skin test (TST) and QuantiFERON-TB-Gold in Tube (QFT) in the diagnosis of latent tuberculosis infection (LTBI) in a prospective community study of contacts of tuberculosis (TB) Jose-Maria García-Garcia 1 , Begoña Gonzalez-Gonzalez 2 , Ana Fernandez-Quiroga 2 , Juan-Jose Palacios 3 , Henar Villar 4 , Jose-Manuel Fernandez-Carreira 5 , Fernando Alvarez-Navascues 1, Marta Garcia-Clemente 1 , Andres Sanchez-Antuña 1 , Jesus Allende 1 , Manuel Martinez-Muñiz 1 , Grupo de Estudio de Contactos de Tuberculosis de Aviles 2 . 1 Neumologia, Hospital San Agustin, Aviles, Asturias, Spain; 2 Atencion Primaria, Area Sanitaria III, Aviles, Asturias, Spain; 3 Unidad de Micobacterias, Hospital Universitario Central de Asturias, Oviedo, Spain; 4 S. Microbiología, Hospital San Agustin, Aviles, Asturias, Spain; 5 U. Investigacion, Hospital San Agustin, Aviles, Asturias, Spain Aim: To evaluate the agreement between TST and QFT in the diagnosis of LTBI. Methods: We performed QFT (according to manufacturer instructions, Cellestis, Australia) and in the same day, after QFT, TST (2 UI of PPD RT23, Mantoux test) in a prospective community study of contacts of patients with TB. If the results of QFT1 and TST1 were – we repeated both (TST2 and QFT2) 10 weeks after. We 925s Abstract printing supported by Chiesi Farmaceutici SpA. Visit Chiesi Farmaceutici SpA. at Stand G.90 Thematic Poster Session Hall 3-53 - 12:50-14:40 T UESDAY, S EPTEMBER 21 ST 2010 defined TST + if induration > or = 5 mm and QFT + if > or = of 0.35 IU/mL. We defined conversion or reversion when the test changed from – to +. Results: TST1 was + in 82/252 (32.5%) and QFT1 in 64/244 (24.6%). TST2 was + in 34/148 (23%) and QFT2 in 20/160 (12.5%). There was concordance between PT1-QFT1 (both + or -) in 197/243 (81.06%) and discordance in 46/243 (18.93%). Concordance in PT2-QFT2 in 110/141 (78.01%) and discordance in 31/141 (21.9%). Overall concordance 79,99%. We had 31 conversions and 3 reversions with TST criteria and 12 conversions and 4 reversions with QFT criteria. In 6 patients we had conversion in TST and QFT. 19 patients had only TST conversion and 2 patients had only QFT conversion. 95 patients had the four tests – (both TST and QFT). Conclusion: In a study of contacts, we found overall concordance between TST and QFT in 80%. We found conversions with both tests (real conversions) although we also found discordance in the conversion between both tests. Both TST and QFT negative supports exposure without infection. Although we found discordances between both test, they are both useful in the diagnosis of LTBI. A grant was provided by Instituto Carlos III (PI 07/90456). As per guidelines, IGRA was indicated in 32 patients but only 2 (6.25%) had the test. 4 patients with abnormal chest X-ray were treated for TB and 17 were offered chemoprophylaxis Conclusion: Data show low use of IGRA test in our DGH. With evidence suggesting this as a cost effective strategy and also helping to screen those, who may not have to undergo chemoprophylaxis treatment with potential toxicities, its use in diagnosing LTBI, should be made widely available. P5004 Study of contacts of tuberculosis. Pilot programme through the coordination of the different assistance levels in the 3rd Sanitary Area of Asturias, Spain Jose-Maria Garcia-Garcia 1 , Ana Fernandez-Quiroga 2 , Begoña Gonzalez-Gonzalez 2 , Victor Amador-Tejon 2 , Jose-Manuel Fernandez-Carreira 3 , Henar Villar 4 , Fernando Alvarez-Navascues 1 , Marta Garcia-Clemente 1 , Andres Sanchez-Antuña 1 , Manuel Martinez-Muñiz 1 , Jesus Allende 1 , Manuel-Angel Villanueva-Montes 1 , Grupo de Estudio de Contactos de Tuberculosis de Aviles 2 . 1 Neumologia, Hospital San Agustin, Aviles, Asturias, Spain; 2 Atencion Primaria, Area Sanitaria III, Aviles, Asturias, Spain; 3 U Investigacion, Hospital San Agustin, Aviles, Asturias, Spain; 4 Microbiologia, Hospital San Agustin, Aviles, Asturias, Spain Aim: To organize the study of contacts of patients with tuberculosis (TB) in a Sanitary Area, and to know the difficulties in the implementation and development of the Study. Methods: We organized a Working Group on TB, with Public Health as epidemiological responsible, Specialized Care as responsible for most of the treatments of TB patients (Respiratory, Internal Medicine, Paediatrics), Microbiology as diagnostic and genetic study responsible, Primary Care in charge of the contacts study and treatment. We had at the beginning a period of formation for the sanitary personnel and a consensus about the study protocol. We collected data about previous BCG, previous TB and treatment, previous tuberculin test and prophylaxis, contact characteristics, symptoms, X-ray, and diagnosis of TB infection (tuberculin test and QuantiFERON). We have opened a website, http://tuberculosisaviles.org/ with access through a user name and password, to introduce the data and therapeutic decisions. We had weekly conferences with programme members to decide what care should be taken in the different people studied. Results: Since 1st of April 2008 we studied 254 contacts of 50 patients with TB. 110 (43.2%) were infected and we started treatment in 73 of them. We found 117 (46%) not infected. 3 contacts had active pulmonary TB (1.1%), 9 (3,5%) contacts did not complete the study and 15 (5,9%) are in follow-up. Conclusion: Our programme permitted the coordination of an important number of sanitary personnel and services involved in the study and was very useful in the study of contacts of TB. A grant was provided by Instituto Carlos III (PI 07/90456). P5005 An audit of diagnosis and chemoprophylaxis treatment of latent tuberculosis Infection (LTBI) in TB contact patients in a district general hospital (DGH) in the United Kingdom (UK) Nawaid Ahmad, Johnny Yin, Dilsher Singh, Ahsan Saleem, Harmesh Moudgil, Julie Beddows, K. Srinivasan. Respiratory Medicine, Princess Royal Hospital, Telford, United Kingdom Background: UK guidelines recommend an algorithm approach to the diagnosis and treatment of patients with LTBI amongst contacts of TB patients. Although Interferon Gamma Release Assay (IGRA) test is recommended in the appropriate subgroup (NICE CG33;2006), adherence to this, is variable. A B Hardy et al (Thorax 2000;65:178-180) have suggested a single step (IGRA test) as more cost effective than the two step (Skin test and IGRA) in diagnosing LTBI. Objective: To audit the use of IGRA in the appropriate group of patients undergoing TB contact testing at our DGH. Method: Retrospective case note analysis of patients undergoing TB contact tracing in our DGH, with particular regard to age, BCG vaccination history, Mantoux test results, IGRA testing and offer of chemoprophylaxis in those diagnosed with LTBI. Results: Table 1 BCG scar present; n=66 (Mantoux =44) BCG scar absent; n=190 (Mantoux=52) Mantoux ≥15mm; n=9 Mantoux≤15mm; n=35 Mantoux ≥6mm; n=23 Mantoux≤6mm; n=29 IGRA n=0, SR n=9 XXXXXXXXXXXX IGRA n=2, SR n=21 BCG n=10 Total = 256 contacts (2007-2009); n = patients, SR = specialist referral. 926s Abstract printing supported by Chiesi Farmaceutici SpA. Visit Chiesi Farmaceutici SpA. at Stand G.90