scholarly journals Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable Mycobacterium tuberculosis

2021 ◽  
Vol 10 (15) ◽  
pp. 3249
Author(s):  
Annelies W. Mesman ◽  
Seung-Hun Baek ◽  
Chuan-Chin Huang ◽  
Young-Mi Kim ◽  
Sang-Nae Cho ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Media ◽  
Multidrug Resistant ◽  
Culture Conditions ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Solid Culture ◽  
Lowenstein Jensen ◽  
Genetic Mechanisms ◽  
Lipid Resuscitation

An estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable Mycobacterium tuberculosis (Mtb) bacilli that do not grow under routine solid culture conditions without the addition of supplementary stimuli. We identified a cohort of TB patients in Lima, Peru, in whom acid-fast bacilli could be detected by sputum smear microscopy, but from whom Mtb could not be grown in standard solid culture media. When we attempted to re-grow Mtb from the frozen sputum samples of these patients, we found that 10 out of 15 could be grown in a glycerol-poor/lipid-rich medium. These fell into the following two groups: a subset that could be regrown in glycerol after “lipid-resuscitation”, and a group that displayed a heritable glycerol-sensitive phenotype that were unable to grow in the presence of this carbon source. Notably, all of the glycerol-sensitive strains were found to be multidrug resistant. Although whole-genome sequencing of the lipid-resuscitated strains identified 20 unique mutations compared to closely related strains, no single genetic lesion could be associated with this phenotype. In summary, we found that lipid-based media effectively fostered the growth of Mtb from a series of sputum smear-positive samples that were not culturable in glycerol-based Lowenstein–Jensen or 7H9 media, which is consistent with Mtb’s known preference for non-glycolytic sources during infection. Analysis of the recovered strains demonstrated that both genetic and non-genetic mechanisms contribute to the observed differential capturability, and suggested that this phenotype may be associated with drug resistance.

scholarly journals Multicenter Study of the Accuracy of the BD MAX Multidrug-resistant Tuberculosis Assay for Detection of Mycobacterium tuberculosis Complex and Mutations Associated With Resistance to Rifampin and Isoniazid

2019 ◽  
Vol 71 (5) ◽  
pp. 1161-1167 ◽  
Author(s):  
Maunank Shah ◽  
Sonia Paradis ◽  
Joshua Betz ◽  
Natalie Beylis ◽  
Renu Bharadwaj ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Resistant Tuberculosis ◽  
Mdr Tb

Abstract Background Tuberculosis (TB) control is hindered by absence of rapid tests to identify Mycobacterium tuberculosis (MTB) and detect isoniazid (INH) and rifampin (RIF) resistance. We evaluated the accuracy of the BD MAX multidrug-resistant (MDR)-TB assay (BD MAX) in South Africa, Uganda, India, and Peru. Methods Outpatient adults with signs/symptoms of pulmonary TB were prospectively enrolled. Sputum smear microscopy and BD MAX were performed on a single raw sputum, which was then processed for culture and phenotypic drug susceptibility testing (DST), BD MAX, and Xpert MTB/RIF (Xpert). Results 1053 participants with presumptive TB were enrolled (47% female; 32% with human immunodeficiency virus). In patients with confirmed TB, BD MAX sensitivity was 93% (262/282 [95% CI, 89–95%]); specificity was 97% (593/610 [96–98%]) among participants with negative cultures on raw sputa. BD MAX sensitivity was 100% (175/175 [98–100%]) for smear-positive samples (fluorescence microscopy), and 81% (87/107 [73–88%]) in smear-negative samples. Among participants with both BD MAX and Xpert, sensitivity was 91% (249/274 [87–94%]) for BD MAX and 90% (246/274 [86–93%]) for Xpert on processed sputa. Sensitivity and specificity for RIF resistance compared with phenotypic DST were 90% (9/10 [60–98%]) and 95% (211/222 [91–97%]), respectively. Sensitivity and specificity for detection of INH resistance were 82% (22/27 [63–92%]) and 100% (205/205 [98–100%]), respectively. Conclusions The BD MAX MDR-TB assay had high sensitivity and specificity for detection of MTB and RIF and INH drug resistance and may be an important tool for rapid detection of TB and MDR-TB globally.

scholarly journals Comparison of different diagnostic modalities for isolation of Mycobacterium Tuberculosis among suspected tuberculous lymphadenitis patients

2023 ◽  
Vol 83 ◽  
Author(s):  
N. Sharif ◽  
D. Ahmed ◽  
R. T. Mahmood ◽  
Z. Qasim ◽  
S. N. Khan ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Communicable Disease ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Primary Objective ◽  
Diagnostic Methods ◽  
Smear Microscopy ◽  
High Morbidity ◽  
Tuberculous Lymphadenitis ◽  
Conventional Methods

Abstract Tuberculosis is a communicable disease with high morbidity and mortality rates in developing countries. The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. The secondary objective is to compare histopathological and microbiological findings in suspected patients with tubercular lymphadenitis. A total of 111 samples (August 2018 to September 2019) of lymph nodes were processed for AFB microscopy, AFB cultures, drug-susceptibility testing (DST), histopathology, and Xpert Mycobacterium Tuberculosis (MTB)/resistance to Rifampin (RIF) assays. Out of 111 lymph node samples, 6 (5.4%) were positive for AFB smear microscopy, 84 (75.6%) were positive for AFB culture, 80 (70.7%) were positive on Gene Xpert, and 102 (91.8%) were indicative of tuberculosis for histopathology studies. Mycobacteria growth indicator tube (MGIT) culture positivity was 84 (75.6%) higher than solid Lowenstein-Jensen (LJ) culture 74 (66.6%). Positive cultures underwent phenotypic DST. Two cases were Multidrug-resistant (MDR) on DST, while three cases were Rifampicin resistant on Gene Xpert. The sensitivity of Genexpert was (62%) against the conventional AFB culture method. The poor performance of conventional lymphadenitis diagnostic methods requires early and accurate diagnostic methodology. Xpert MTB/RIF test can help in the treatment of multidrug-resistant TB cases. Nonetheless, rapid and conventional methods should be used for complete isolation of Mycobacterium tuberculosis.

scholarly journals Sensitivitas Metode Pemeriksaan Mikroskopis Fluorokrom dan Ziehl-Neelsen untuk Deteksi Mycobacterium tuberculosis pada Sputum

2019 ◽  
Vol 1 (2) ◽  
pp. 56
Author(s):  
BETTY SURYAWATI ◽  
LELI SAPTAWATI ◽  
ASTARI FEBYANE PUTRI ◽  
JATU APHRIDASARI
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Early Detection ◽  
Pulmonary Tuberculosis ◽  
Sensitivity And Specificity ◽  
Smear Microscopy ◽  
Microbiology Laboratory ◽  
Fluorochrome Staining ◽  
Primary Screening ◽  
Lowenstein Jensen ◽  
Tuberculosis Diagnosis

<p class="AbstractNormal"><em>Background: Detection of fast acid bacteria (FAB) using smear microscopy is used as a primary screening for tuberculosis diagnosis. Previous studies have shown that fluorochrome </em>(<em>Auroamine-rhodamine</em>) <em>staining showed better sensitivity compared to Ziehl-Neelsen (ZN) method in the detection of FAB in sputum. However this method has not been recommended for routine use including in Indonesia. This study aimed to evaluate the sensitivity and specificity of fluorochrome compared to ZN to detect FAB in patient’s sputum.</em><em></em></p><p class="AbstractNormal"><em>Methods: </em><em>This study analyzed 60 sputum samples from patients with tuberculosis and suspected pulmonary tuberculosis. Samples were obtained consecutively from microbiology laboratory</em><em> Moewardi Hospital, Indonesia. Each sample was examined using ZN and fluorochrome staining and cultured in Lowenstein-Jensen (LJ) medium.</em><em> Data were analyzed using sensitivity and spesificity tests.</em></p><p class="AbstractNormal"><em>Results: ZN staining detected FAB in 12 samples (10%), while fluorochrome detected FAB in 17 samples (28%). The sensitivity and specificity of ZN staining were 70% and 90% while these for fluorochrome were 90% and 84%. </em><em></em></p><p class="AbstractNormal"><em>Conclusions: The sensitivity of fluorochrome staining is better compared to ZN staining. This method can be recommended for early detection of tuberculosis.</em><em></em></p><p class="AbstractNormal"><em> </em></p>

Prevalence and Distribution of Multidrug-Resistant Mutations in Mycobacterium tuberculosis in Tanzania

2019 ◽  
Vol 1 (1) ◽  
pp. 15-22
Author(s):  
John C. Mgogwe ◽  
Hadija H. Semvua ◽  
Oliva Safari ◽  
Gibson E. Kapanda ◽  
Balthazar M. Nyombi ◽  
...  
Keyword(s):  
Gene Mutations ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Sputum Smear ◽  
Sensitivity Testing ◽  
Cross Sectional ◽  
Important Approach ◽  
Mdr Tb ◽  
Frequent Mutations ◽  
Lowenstein Jensen

Background: Molecular identification of mutations resulting in multidrug-resistant tuberculosis (MDR-TB) is an important approach for improving understanding of MDR-TB epidemiology and planning for appropriate interventions. We aimed to estimate the prevalence and distribution of mutations causing MDR-TB as well as determine the gene distribution among patients previously treated for TB. Methods: This was a cross-sectional, hospital-based study conducted from April 2017 to October 2018 at Kibong’oto Infectious Diseases Hospital (KIDH). KIDH is the national MDR-TB referral hospital. Participants were patients presumptively diagnosed with MDR-TB and referred to KIDH from district and regional hospitals across Tanzania. Sputum samples were collected and analysed using the Xpert MTB/RIF assay, direct sputum smear fluorescence microscopy, culture on Lowenstein-Jensen medium, and line probe assay using the GenoType MTBDRplus VER 2.0 system. Demographic information and mutation frequencies were reported as counts and percentages and analysed using descriptive statistics. Results: A total of 208 (69.3%) participants had rpoB gene mutations conferring resistance to only rifampicin; 92 (30.7%) had rpoB, katG, and inhA mutations conferring resistance to rifampicin and isoniazid; 78 (26%) had rpoB and katG mutations conferring resistance to rifampicin and isoniazid; and 14 (4.7%) had rpoB and inhA mutations conferring resistance to rifampicin and isoniazid. Conclusion: The mutation prevalences identified in this study indicate the most frequent mutations were the S531L mutation of the rpoB gene, the S315T1 mutation of the katG gene, and the S315T mutation in the promoter region of the inhA gene. To control the emergence and spread of MDR-TB, drug sensitivity testing must be carried for GeneXpert-confirmed TB patients prior to initiating second-line anti-TB regimens.

scholarly journals NILAI DIAGNOSTIK ANTIGEN TB DENGAN RAPID TEST DEVICE (TB AG) UNTUK TUBERKULOSIS PARU

2016 ◽  
Vol 18 (3) ◽  
pp. 161
Author(s):  
Sri Kartika Sari ◽  
Aryati Aryati
Keyword(s):  
Pulmonary Tuberculosis ◽  
Sensitivity And Specificity ◽  
Rapid Test ◽  
Diagnostic Value ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Test Device ◽  
Tuberculosis Patients ◽  
Suspected Tuberculosis ◽  
Lowenstein Jensen

In Indonesia, the diagnosis of pulmonary tuberculosis relies primarily on an identification of acid-fast bacilli on sputum smears. However, microscopic device has several limitations. The sensitivity of microscopic examination is variable. The quality of smear microscopic results is heavily depend on the workload, and the skill of the technician’s reading the slide. TB antigen rapid test device (TB Ag) is fast, easy and does not either need skillness of the operator. The kit detects specific secreted antigen M.tuberculosis coded by: RD (Region of Difference) 1, RD2 and RD3. These RD1−3 were found deleted from BCG (Bacille Calmette-Guerine) vaccine strain. In the present study, the diagnostic value of TB Ag was assessed. Sputum samples were examined from 59 suspected tuberculosis patients and 22 non tuberculosis patients. The samples of the suspected tuberculosis patients were collected as three consecutive sputum specimens (spot, morning, spot). The total 199 specimens were examined by sputum smear microscopy and TB Ag. M.tuberculosis culture by using Lowenstein Jensen media, which was used as a gold standard. The sensitivity and specificity of microscopic sputum smear were 83.8% (95% CI: 70.0–89.4) and 96.3% (95% CI: 89.8–98.7), respectively. While, the sensitivity and specificity of TB Ag were 72.6% (95% CI: 63.9–79.9) and 90.9% (95% CI: 72.2–97.5), respectively. The concordance between microscopic sputum smear and TB Ag was 70.8%. TB Ag can be considered as a new diagnostic tool for the diagnosis of pulmonary tuberculosis, especially at the health services where there is no expert technician available for microscopic sputum smear examination.

scholarly journals The Mono-Prep system increases the detection rate of sputum smear microscopy for diagnosing tuberculosis

2018 ◽  
Vol 46 (12) ◽  
pp. 5137-5142
Author(s):  
Jiannan Wu ◽  
Chengcheng Kong ◽  
Fengmin Huo ◽  
Qian Liang ◽  
Yifeng Ma ◽  
...  
Keyword(s):  
Detection Rate ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Sputum Smear Microscopy ◽  
Solid Culture ◽  
Direct Smear ◽  
Alternative Method ◽  
Lower Specificity ◽  
Positive Rate

Objective Direct sputum smear microscopy (DSSM) has a low detection rate. This study investigated whether an alternative method called Mono-Prep smear microscopy (MPSM) can enhance the diagnosis of tuberculosis in tuberculosis laboratories that perform direct smear microscopy in China. Methods A total of 117 sputum samples were collected from outpatients who attended Beijing Chest Hospital. DSSM, MPSM, solid culture, and Xpert MTB/RIF were performed on the samples. Results The positive rates of DSSM, MPSM, solid culture, and Xpert MTB/RIF were 27.4% (32/117), 40.2% (47/117), 35.9% (42/117), and 52.1% (61/117), respectively. MPSM could detect 15 more cases of tuberculosis compared with DSSM (47 vs 32) among 117 sputum samples. This represented a significantly higher positive rate and sensitivity of MPSM compared with DSSM. However, MPSM appeared to have a lower specificity (81.3%) compared with DSSM (90.7%) with the solid culture used as a standard. Conclusion Use of MPSM can increase the number of positive sputum samples, but it still needs improvement.

scholarly journals The T2 Mycobacterium tuberculosis Genotype, Predominant in Kampala, Uganda, Shows Negative Correlation with Antituberculosis Drug Resistance

2014 ◽  
Vol 58 (7) ◽  
pp. 3853-3859 ◽  
Author(s):  
Deus Lukoye ◽  
Fred A. Katabazi ◽  
Kenneth Musisi ◽  
David P. Kateete ◽  
Benon B. Asiimwe ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Epidemiological Studies ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Sputum Smear ◽  
Hiv Status ◽  
Content Type ◽  
Phylogenetic Lineages

ABSTRACTSurveillance of the circulatingMycobacterium tuberculosiscomplex (MTC) strains in a given locality is important for understanding tuberculosis (TB) epidemiology. We performed molecular epidemiological studies on sputum smear-positive isolates that were collected for anti-TB drug resistance surveillance to establish the variability of MTC lineages with anti-TB drug resistance and HIV infection. Spoligotyping was performed to determine MTC phylogenetic lineages. We compared patients' MTC lineages with drug susceptibility testing (DST) patterns and HIV serostatus. Out of the 533 isolates, 497 (93.2%) had complete DST, PCR, and spoligotyping results while 484 (90.1%) participants had results for HIV testing. Overall, the frequency of any resistance was 75/497 (15.1%), highest among the LAM (34.4%; 95% confidence interval [CI], 18.5 to 53.2) and lowest among the T2 (11.5%; 95% CI, 7.6 to 16.3) family members. By multivariate analysis, LAM (adjusted odds ratio [ORadj], 5.0; 95% CI, 2.0 to 11.9;P< 0.001) and CAS (ORadj, 2.9; 95% CI, 1.4.0 to 6.3;P= 0.006) families were more likely to show any resistance than was T2. All other MTC lineages combined were more likely to be resistant to any of the anti-TB drugs than were the T2 strains (ORadj, 1.7; 95% CI, 1.0 to 2.9;P= 0.040). There were no significant associations between multidrug resistance and MTC lineages, but numbers of multidrug-resistant TB strains were small. No association was established between MTC lineages and HIV status. In conclusion, the T2 MTC lineage negatively correlates with anti-TB drug resistance, which might partly explain the reported low levels of anti-TB drug resistance in Kampala, Uganda. Patients' HIV status plays no role with respect to the MTC lineage distribution.

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