Detection of tuberculous mycobacteria in sputum and intensity of bacillary excretion in tuberculosis patients with various HIV status

Goal of the study: to investigate the informativeness of tuberculous mycobacteria detection by various techniques and to analyze the intensity of bacillary excretion in tuberculosis patients depending on their HIV status. Materials and methods. 360 HIV patients were examined due to suspected tuberculosis. The diagnostic procedure included two consecutive sputum test for acid fast bacilli (AFB)/tuberculous mycobacteria by fluorescent microscopy, molecular genetic tests and culture on liquid and solid media. Tuberculosis was diagnosed in 178 (49.4%) patients. The comparison group included 354 tuberculosis patients with negative HIV status including 113 patients who underwent the same diagnostic procedure as those suffering from TB/HIV co-infection. results of the study. AFB were more often detected by fluorescent microscopy in HIV negative tuberculosis patients compared to those with TB/HIV co-infection (32.8 and 24.2% respectively), and the sensitivity of the other testing methods did not differ. Scarce bacillary excretion was confidently more often registered in those suffering from TB/HIV co-infection and less frequently they had massive bacillary excretion detected by culture on solid media compared to HIV negative tuberculosis patients. The complete diagnostic procedure demonstrated better sensitivity in those with TB/HIV co-infection compared to tuberculosis patients with negative HIV status (87.1 and 71.2%). © 2018 New Terra Publishing House. All Rights Reserved

Detection of tuberculous mycobacteria in sputum and intensity of bacillary excretion in tuberculosis patients with various HIV status

Goal of the study: to investigate the informativeness of tuberculous mycobacteria detection by various techniques and to analyze the intensity of bacillary excretion in tuberculosis patients depending on their HIV status. Materials and methods. 360 HIV patients were examined due to suspected tuberculosis. The diagnostic procedure included two consecutive sputum test for acid fast bacilli (AFB)/tuberculous mycobacteria by fluorescent microscopy, molecular genetic tests and culture on liquid and solid media. Tuberculosis was diagnosed in 178 (49.4%) patients. The comparison group included 354 tuberculosis patients with negative HIV status including 113 patients who underwent the same diagnostic procedure as those suffering from TB/HIV co-infection. results of the study. AFB were more often detected by fluorescent microscopy in HIV negative tuberculosis patients compared to those with TB/HIV co-infection (32.8 and 24.2% respectively), and the sensitivity of the other testing methods did not differ. Scarce bacillary excretion was confidently more often registered in those suffering from TB/HIV co-infection and less frequently they had massive bacillary excretion detected by culture on solid media compared to HIV negative tuberculosis patients. The complete diagnostic procedure demonstrated better sensitivity in those with TB/HIV co-infection compared to tuberculosis patients with negative HIV status (87.1 and 71.2%). © 2018 New Terra Publishing House. All Rights Reserved

Выявление микобактерий туберкулеза в мокроте у больных ВИЧ-инфекцией при использовании современного алгоритма этиологической диагностики заболевания

The objective. To establish the informativity of using phenotypic and molecular genetic methods of detection of mycobacterium tuberculosis (МВТ) from sputum to diagnose tuberculosis in patients with HIV infection. Patients and methods. 360 HIV-infected individuals with suspected tuberculosis were examined. A diagnostic algorithm included two tests of sputum for AFB/ МВТ by fluorescence microscopy, detection of МВТ DNA in real-time PCR and inoculation of liquid and solid media. Results. Tuberculosis was diagnosed in 49.4% of the examined patients. The greatest sensitivity and accuracy of МВТ detection was demonstrated by molecular genetic methods (71.3% and 0.85, respectively), which even exceeded tire results of culture diagnosis (sensitivity - 58.4 and 60.7%, accuracy - 0.79 and 0.81 for inoculation of liquid and dense media, respectively). An extremely low sensitivity was demonstrated by fluorescence microscopy (24.2%). No relation between the frequency of МВТ detection and the number of CD4+-lymphocytes was found in any method but for fluorescence microscopy showed a tendency of a better effectiveness in patients with profound immunodeficiency (in CD4 less than 100 cell/pl). Repeat analysis of inoculation of liquid media and detection of МВТ DNA (MGM) enhanced the diagnostic value of each method by 8.6 and 7.1%, respectively. In 73.5% of patients, tuberculosis was diagnosed within 2-3 days from the moment of turning for medical help. Conclusion. In general, an examination algorithm for detection of МВТ demonstrated its sensitivity in patients with co-infection (HIV/ТВ) in 87.1%. If molecular genetic methods can be used for rapid detection of a causative agent fluorescence microscopy looses its relevance for diagnosing tuberculosis.Цель. Установить информативность использования фенотипических и молекулярно-генетических методов детекции микобактерии туберкулезис (МВТ) из мокроты для диагностики туберкулеза у больных ВИЧ-инфекцией. Пациенты и методы. Обследование выполнено 360 ВИЧ-инфицированным с подозрением на туберкулез. Диагностический алгоритм включал двукратное исследование мокроты на КУМ/МБТ люминесцентной микроскопией, выявлением ДНК МВТ с помощью полимеразной цепной реакции реального времени и посевом на жидкие и плотные питательные среды. Результаты. Туберкулез установлен 49,4% пациентам от числа обследованных. Наибольшую чувствительность и точность в выявлении МБТ показали молекулярно-генетические методы (71,3% и 0,85 соответственно), которые даже превысили результаты культуральной диагностики (чувствительность - 58,4 и 60,7%, точность - 0,79 и 0,81 для посева на жидкие и плотные среды соответственно). Крайне низкую чувствительность показала люминесцентная микроскопия (24,2%). Не отмечено связи между частотой выявления МБТ и количеством СД4+-лимфоцитов ни по одному из методов, однако для люминесцентной микроскопии отмечена тенденция лучшей результативности у больных с глубоким иммунодефицитом (при CD4 менее 100 клеток/мкл). Повторный анализ посева на жидкие среды и выявления ДНК МБТ (МГМ) усилили диагностическую ценность каждого из методов на 8,6 и 7,1% соответственно. У 73,5% больных диагноз туберкулеза был установлен в течение 2-3 дней от момента обращения за помощью. Заключение. В целом алгоритм обследования для выявления МБТ продемонстрировал чувствительность у больных ко-инфекцией (ВИЧ/ТБ) 87,1%. При возможности использовать быстрые молекулярно-генетические методы детекции возбудителя люминесцентная микроскопия теряет свою значимость в диагностике туберкулеза

The spectrum of primary drug resistance of Mycobacterium tuberculosis in patients with tuberculosis in relation to human immunodeficiency virus status

Aim. To estimate the detection rate and spectrum of primary drug resistance of Mycobacterium tuberculosis (MBT) in patients with tuberculosis (TB) in relation to their human immunodeficiency virus (HIV) status in a region with high HIV infection rates (the Perm Territory) and to compare of drug-resistant MBT (DR-MBT) in patients with HIV/TB co-infection, by using phenotypic and molecular genetic testing (MGT) methods. Subjects and methods. The results of sputum bacteriological examination were analyzed in 178 HIV-infected patients and 354 non-HIV-infected individuals with a TB diagnosis made in the period July 1, 2014 to August 1, 2015. The diagnostic algorithm for all patients involved a duplicate sputum test for MBT by two techniques: fluorescence microscopy (FM) and inoculation into the Levenstein-Jensen dense culture medium. In patients with HIV/TB, the bacteriological examination was complemented with two more methods: detection of MBT DNA by a real-time polymerase chain reaction assay using the AmpliTube-RV system (Synthol, Russia); and inoculation into the Middlebrook liquid nutrient medium, by applying the automated BACTEC MGIT 960 system. Results. In patients with HIV/TB, the sensitivity of FM proved to be lower than in those with TB (24.2 and 32.8%, respectively; p0.05). The primary drug resistance of MBT in patients with HIV-TB was higher than that in HIV-negative individuals (60.2 and 41.6%, respectively; p<0.05). The phenotypic method (inoculation into the Levenstein-Jensen culture medium) and MGT revealed their agreement for the resistance of MBT to rifampicin (the most clinically significant drug in the choice of treatment policy) in 88.5% of the patients with HIV/TB. Conclusion. In patients with HIV/TB, the sensitivity of FM for detecting acid-resistant mycobacteria was lower than in those with TB and that of inoculations into the dense medium was comparable regardless of HIV status

Выявление микобактерий туберкулеза в мокроте у больных ВИЧ-инфекцией при использовании современного алгоритма этиологической диагностики заболевания

The objective. To establish the informativity of using phenotypic and molecular genetic methods of detection of mycobacterium tuberculosis (МВТ) from sputum to diagnose tuberculosis in patients with HIV infection. Patients and methods. 360 HIV-infected individuals with suspected tuberculosis were examined. A diagnostic algorithm included two tests of sputum for AFB/ МВТ by fluorescence microscopy, detection of МВТ DNA in real-time PCR and inoculation of liquid and solid media. Results. Tuberculosis was diagnosed in 49.4% of the examined patients. The greatest sensitivity and accuracy of МВТ detection was demonstrated by molecular genetic methods (71.3% and 0.85, respectively), which even exceeded tire results of culture diagnosis (sensitivity - 58.4 and 60.7%, accuracy - 0.79 and 0.81 for inoculation of liquid and dense media, respectively). An extremely low sensitivity was demonstrated by fluorescence microscopy (24.2%). No relation between the frequency of МВТ detection and the number of CD4+-lymphocytes was found in any method but for fluorescence microscopy showed a tendency of a better effectiveness in patients with profound immunodeficiency (in CD4 less than 100 cell/pl). Repeat analysis of inoculation of liquid media and detection of МВТ DNA (MGM) enhanced the diagnostic value of each method by 8.6 and 7.1%, respectively. In 73.5% of patients, tuberculosis was diagnosed within 2-3 days from the moment of turning for medical help. Conclusion. In general, an examination algorithm for detection of МВТ demonstrated its sensitivity in patients with co-infection (HIV/ТВ) in 87.1%. If molecular genetic methods can be used for rapid detection of a causative agent fluorescence microscopy looses its relevance for diagnosing tuberculosis.Цель. Установить информативность использования фенотипических и молекулярно-генетических методов детекции микобактерии туберкулезис (МВТ) из мокроты для диагностики туберкулеза у больных ВИЧ-инфекцией. Пациенты и методы. Обследование выполнено 360 ВИЧ-инфицированным с подозрением на туберкулез. Диагностический алгоритм включал двукратное исследование мокроты на КУМ/МБТ люминесцентной микроскопией, выявлением ДНК МВТ с помощью полимеразной цепной реакции реального времени и посевом на жидкие и плотные питательные среды. Результаты. Туберкулез установлен 49,4% пациентам от числа обследованных. Наибольшую чувствительность и точность в выявлении МБТ показали молекулярно-генетические методы (71,3% и 0,85 соответственно), которые даже превысили результаты культуральной диагностики (чувствительность - 58,4 и 60,7%, точность - 0,79 и 0,81 для посева на жидкие и плотные среды соответственно). Крайне низкую чувствительность показала люминесцентная микроскопия (24,2%). Не отмечено связи между частотой выявления МБТ и количеством СД4+-лимфоцитов ни по одному из методов, однако для люминесцентной микроскопии отмечена тенденция лучшей результативности у больных с глубоким иммунодефицитом (при CD4 менее 100 клеток/мкл). Повторный анализ посева на жидкие среды и выявления ДНК МБТ (МГМ) усилили диагностическую ценность каждого из методов на 8,6 и 7,1% соответственно. У 73,5% больных диагноз туберкулеза был установлен в течение 2-3 дней от момента обращения за помощью. Заключение. В целом алгоритм обследования для выявления МБТ продемонстрировал чувствительность у больных ко-инфекцией (ВИЧ/ТБ) 87,1%. При возможности использовать быстрые молекулярно-генетические методы детекции возбудителя люминесцентная микроскопия теряет свою значимость в диагностике туберкулеза

The spectrum of primary drug resistance of Mycobacterium tuberculosis in patients with tuberculosis in relation to human immunodeficiency virus status

Aim. To estimate the detection rate and spectrum of primary drug resistance of Mycobacterium tuberculosis (MBT) in patients with tuberculosis (TB) in relation to their human immunodeficiency virus (HIV) status in a region with high HIV infection rates (the Perm Territory) and to compare of drug-resistant MBT (DR-MBT) in patients with HIV/TB co-infection, by using phenotypic and molecular genetic testing (MGT) methods. Subjects and methods. The results of sputum bacteriological examination were analyzed in 178 HIV-infected patients and 354 non-HIV-infected individuals with a TB diagnosis made in the period July 1, 2014 to August 1, 2015. The diagnostic algorithm for all patients involved a duplicate sputum test for MBT by two techniques: fluorescence microscopy (FM) and inoculation into the Levenstein-Jensen dense culture medium. In patients with HIV/TB, the bacteriological examination was complemented with two more methods: detection of MBT DNA by a real-time polymerase chain reaction assay using the AmpliTube-RV system (Synthol, Russia); and inoculation into the Middlebrook liquid nutrient medium, by applying the automated BACTEC MGIT 960 system. Results. In patients with HIV/TB, the sensitivity of FM proved to be lower than in those with TB (24.2 and 32.8%, respectively; p0.05). The primary drug resistance of MBT in patients with HIV-TB was higher than that in HIV-negative individuals (60.2 and 41.6%, respectively; p<0.05). The phenotypic method (inoculation into the Levenstein-Jensen culture medium) and MGT revealed their agreement for the resistance of MBT to rifampicin (the most clinically significant drug in the choice of treatment policy) in 88.5% of the patients with HIV/TB. Conclusion. In patients with HIV/TB, the sensitivity of FM for detecting acid-resistant mycobacteria was lower than in those with TB and that of inoculations into the dense medium was comparable regardless of HIV status

Healthcare visits of HIV-infected persons before and after tuberculosis diagnosis and contamination of the environment of health-care facilities with mycobacterium tuberculosis

The objective. To study healthcare visits of HIV-infected persons before and after they were diagnosed with tuberculosis and contamination of healthcare facilities (HF) with mycobacterium tuberculosis. Materials and methods. We studied medical records of 169 HIV-infected patients who fell ill with tuberculosis. The frequency of visits of HIV-infected patients to various HF before and after tuberculosis diagnosis was determined. Molecular-genetic examination of 316 smears was performed for the presence of mycobacterium tuberculosis (MBT) DNA, taken from the working environment of four HF (AIDS Centre clinic, infectious inpatient hospital for HIV-infected persons, tuberculosis inpatient clinic, local general-duty polyclinic). Results. As has been found, before and after tuberculosis diagnosis HIV-infected persons most commonly apply for medical aid to the AIDS Centre polyclinic, more rarely to other HF. Tuberculosis in HIV-infected persons is most commonly diagnosed in the AIDS Centre polyclinic, more rarely in general-duty polyclinic, department of the infectious hospital for HIV-infected patients, somatic and surgical inpatient units. According to molecular-genetic examination of environmental smear sampling in various healthcare facilities, the maximal contamination with MBT DNA was found in the AIDS Centre polyclinic, in a less degree in the tuberculosis inpatient hospital and the infectious inpatient hospital for HIV-infected persons. In the general-duty polyclinic, no mycobacterium tuberculosis DNA was isolated. Conclusion. Predominant visits of HIV-infected persons before and after tuberculosis diagnosis to the AIDS Centre polyclinic and a higher level of contamination of healthcare environment in this facility with MBT DNA show that in the settings of a specialised polyclinic there are conditions for nosocomial infection with tuberculosis bacteria

Healthcare visits of HIV-infected persons before and after tuberculosis diagnosis and contamination of the environment of health-care facilities with mycobacterium tuberculosis

The objective. To study healthcare visits of HIV-infected persons before and after they were diagnosed with tuberculosis and contamination of healthcare facilities (HF) with mycobacterium tuberculosis. Materials and methods. We studied medical records of 169 HIV-infected patients who fell ill with tuberculosis. The frequency of visits of HIV-infected patients to various HF before and after tuberculosis diagnosis was determined. Molecular-genetic examination of 316 smears was performed for the presence of mycobacterium tuberculosis (MBT) DNA, taken from the working environment of four HF (AIDS Centre clinic, infectious inpatient hospital for HIV-infected persons, tuberculosis inpatient clinic, local general-duty polyclinic). Results. As has been found, before and after tuberculosis diagnosis HIV-infected persons most commonly apply for medical aid to the AIDS Centre polyclinic, more rarely to other HF. Tuberculosis in HIV-infected persons is most commonly diagnosed in the AIDS Centre polyclinic, more rarely in general-duty polyclinic, department of the infectious hospital for HIV-infected patients, somatic and surgical inpatient units. According to molecular-genetic examination of environmental smear sampling in various healthcare facilities, the maximal contamination with MBT DNA was found in the AIDS Centre polyclinic, in a less degree in the tuberculosis inpatient hospital and the infectious inpatient hospital for HIV-infected persons. In the general-duty polyclinic, no mycobacterium tuberculosis DNA was isolated. Conclusion. Predominant visits of HIV-infected persons before and after tuberculosis diagnosis to the AIDS Centre polyclinic and a higher level of contamination of healthcare environment in this facility with MBT DNA show that in the settings of a specialised polyclinic there are conditions for nosocomial infection with tuberculosis bacteria