1. health and fitness
2. disease

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
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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
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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
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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
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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.
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