Increased sensitivity of Mycobacterium tuberculosis Cobas Amplicor PCR following brief incubation of tissue samples on Lowenstein-Jensen substrate

Apmis ◽  
2003 ◽  
Vol 111 (12) ◽  
pp. 1114-1116 ◽  
Author(s):  
MARIANA CRISTEA FERNSTROM ◽  
LENA DAHLGREN ◽  
MARIA RANBY ◽  
ARNE FORSGREN ◽  
BJORN PETRINI
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Tissue Samples ◽  
Lowenstein Jensen ◽  
Increased Sensitivity

Determination some Immunological Aspects in Pulmonary Tuberculosis Patients in Qadisiyah Province

2018 ◽  
Vol 9 (2) ◽  
Author(s):  
Syoof Khowman Alramahy ◽  
Akram Hadi Hamza
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Statistical Difference ◽  
Positive Culture ◽  
Control Group ◽  
Tuberculosis Patients ◽  
Significant Difference ◽  
Lowenstein Jensen ◽  
The Mean ◽  
Igg Level

This study was carried out to study of some immunological aspects among the pulmonary Tuberculosis patients infected with causative agent, Mycobacterium tuberculosis. A Total of 200 sputum samples were collected from patients attending the consultant Clinic for Chest and Respiratory disease center, Diwaniya. Control group (No=15) also included. According to acid fast stain of sputum, the patients were classified as positive (No=91,45.5%) and negative (No=109,54.5, Lowenstein Jensen medium used for the cultivation of samples, on which 70% of sputum samples where positive culture for this microorganism. The grown microorganism were identified as M. tuberculosis, based on positive A.F.B, Niacin producers ,negative for catlase at 68c. The mean IgG level was l184.053±76.684 mg/100 ml in tuberculosis group compared with 1016.533 ± 44.882 mg/100ml in control group, rendering the statistical difference significant. For IgA and IgM levels, they were at mean of 315.880±38.552 mg/100 ml and 119.527±8.464 mg/100 ml in control group compared with 396.358±38.776 mg/100 ml and 134.207±11.696 mg/100 ml in patients group respectively with significant difference

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 Bovine tuberculosis in Rwanda: Prevalence and economic impact evaluation by meat inspection at Société des Abattoirs de Nyabugogo-Nyabugogo Abattoir, Kigali

2014 ◽  
Vol 85 (1) ◽  
Author(s):  
Gervais Habarugira ◽  
Joseph Rukelibuga ◽  
Mark O. Nanyingi ◽  
Borden Mushonga
Keyword(s):  
Economic Impact ◽  
Bovine Tuberculosis ◽  
Meat Inspection ◽  
Control Measures ◽  
Meat Industry ◽  
Biochemical Tests ◽  
Tissue Samples ◽  
Public Health Burden ◽  
Significant Public Health ◽  
Lowenstein Jensen

Despite the significant public health burden of bovine tuberculosis (bTB) in Rwanda, the prevalence of bTB is poorly documented. This study was conducted to estimate the prevalence of bTB in cattle using gross examination of granulomatous lesions, to identify mycobacteria species in suspected samples, and to evaluate the economic impact of meat condemnation based on bTB-like lesions in the meat industry in Rwanda. Routine meat inspection was conducted at Société des Abattoirs de Nyabugogo (SABAN)-Nyabugogo Abattoir. Tissue samples including 31 lymph nodes, 3 lungs and 2 livers were obtained from cattle of different ages with gross tuberculous lesions. Mycobacterium bovis was identified using microscopy with Kinyoun staining and isolation of mycobacterial species in culture on Löwenstein–Jensen and Colestos media, further identified using biochemical tests. Our findings, based on culture and postmortem results, show that the prevalence of bTB is 0.5%(0.587*148/16753), with an overall gross tuberculous lesion prevalence of 0.9% (148/16753). The presence of lesions were higher in cattle aged 2 years and older (1.6% vs. 0.6%, p < 0.05) and higher in females than in males (1.4% vs. 0.6%, p < 0.05). Of the 36 samples tested, 26 (72.2%) were positive by microscopic examination with Kinyoun staining while M. bovis was culture-confirmed in 21 (58.7%) cases. Bovine tuberculosis caused condemnation of 1683.5 kg of meat, resulting in an estimated loss of $4810. Our findings indicate that the prevalence of bTB in Rwanda is significant, and that bTB is a major cause of meat condemnation requiring continued implementation of surveillance and control measures. Furthermore, the results from this study also show important variations in sensitivity of the different tests that were used to determine the prevalence of bTB in cattle in Rwanda.

scholarly journals Evaluation of the Xpert MTB/RIF Performance on Tissues: Potential Impact on Airborne Infection Isolation at a Tertiary Cancer Care Center

2018 ◽  
Vol 39 (4) ◽  
pp. 462-466 ◽  
Author(s):  
Tracy McMillen ◽  
Shauna C. Usiak ◽  
Liang Hua Chen ◽  
Luz Gomez ◽  
Peter Ntiamoah ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Limit Of Detection ◽  
Tertiary Care ◽  
Ffpe Tissue ◽  
Cancer Center ◽  
Oncology Patients ◽  
Tissue Samples ◽  
Airborne Infection ◽  
Potential Impact ◽  
Tissue Specimens

OBJECTIVESIn this study, we sought to evaluate the performance of the Xpert MTB/RIF (Cepheid) assay for the detection of Mycobacterium tuberculosis (MTB) complex DNA on fresh and formalin-fixed, paraffin-embedded (FFPE) tissue specimens from oncology patients in an area with a low prevalence of tuberculosis. We also aimed to retrospectively assess the potential impact of Xpert MTB/RIF on the duration of airborne infection isolation (AII).SETTINGA 473-bed, tertiary-care cancer center in New York City.DESIGNA total of 203 tissue samples (101 FFPE and 102 fresh) were tested using Xpert MTB/RIF, including 133 pulmonary tissue samples (65.5%) and 70 extrapulmonary tissue samples (34.5%). Acid-fast bacilli (AFB) culture was used as the diagnostic gold standard. The limit of detection (LOD) and reproducibility were also evaluated for both samples types using contrived specimens. The potential impact of the Xpert MTB PCR assay on tissue samples from AII patients on AII duration was retrospectively assessed.RESULTSUsing the Xpert MTB/RIF for fresh tissue specimens, the sensitivity was 50% (95% CI, 1.3%–98.7%) and the specificity was 99% (95% CI, 94.5%–99.9%). For FFPE tissue specimens, the sensitivity was 100% (95% CI, 63.1%–100%) and the specificity was 98.3% (95% CI, 95.5%–100%. The LOD was 103 colony-forming units (CFU)/mL for both fresh and FFPE tissue specimens, and the Xpert MTB/RIF was 100% reproducible at concentrations 10 times that of the LOD. With an expected turnaround time of 24 hours, the Xpert MTB PCR could decrease the duration of AII from a median of 8 days to a median of 1 day.CONCLUSIONSThe Xpert MTB/RIF assay offers a valid option for ruling out Mycobacterium tuberculosis complex (MTBC) on tissue samples from oncology patients and for minimizing AII resource utilization.Infect Control Hosp Epidemiol 2018;39:462–466

scholarly journals Multicenter Evaluation of the BACTEC MGIT 960 System for Recovery of Mycobacteria

1999 ◽  
Vol 37 (3) ◽  
pp. 748-752 ◽  
Author(s):  
Bruce A. Hanna ◽  
Adeleh Ebrahimzadeh ◽  
L. Bruce Elliott ◽  
Margie A. Morgan ◽  
Susan M. Novak ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Liquid Medium ◽  
Mycobacterium Avium ◽  
Culture Systems ◽  
Solid Media ◽  
Lowenstein Jensen ◽  
The Mean

We evaluated the BACTEC MGIT 960 system, which is a fully automated, noninvasive system for the growth and detection of mycobacteria with a capacity to incubate and continuously monitor 960 7-ml culture tubes. We studied 3,330 specimens, 2,210 respiratory and 1,120 nonrespiratory specimens, collected from 2,346 patients treated at six sites. Processed specimens were inoculated into the BACTEC MGIT 960 and BACTEC 460 TB systems, as well as onto Lowenstein-Jensen slants and Middlebrook 7H11/7H11 selective plates. From all culture systems, a total of 362 isolates of mycobacteria were recovered; these were recovered from 353 specimens collected from 247 patients. The greatest number of isolates of mycobacteria (289, or 80% of the 362 isolates) was recovered with the BACTEC MGIT 960, followed by the BACTEC 460 TB (271, or 75%) and solid media (250, or 69%). From all culture systems a total of 132 isolates of Mycobacterium tuberculosiscomplex were recovered. The greatest number of isolates of M. tuberculosis complex was recovered when liquid medium was combined with conventional solid media; the number recovered with BACTEC 460 TB plus solid media was 128 (97%), that recovered with BACTEC MGIT 960 plus solid media was 121 (92%), that recovered with BACTEC 460 TB was 119 (90%) and that recovered with all solid media combined was 105 (79%). The recovery with BACTEC MGIT 960 alone was 102 (77%). The mean times to detection (TTD) for M. tuberculosis complex were 14.4 days for BACTEC MGIT, 15.2 days for BACTEC 460 TB, and 24.1 days for solid media. The numbers of isolates of Mycobacterium avium complex (MAC) recovered were 172 (100%) for all systems, 147 (85%) for BACTEC MGIT 960, 123 (72%) for BACTEC 460 TB, and 106 (62%) for all solid media combined. The TTD for MAC in each system were 10.0 days for BACTEC MGIT 960, 10.4 days for BACTEC 460 TB, and 25.9 days for solid media. Breakthrough contamination rates (percentages of isolates) for each of the systems were 8.1% for BACTEC MGIT 960, 4.9% for BACTEC 460 TB, and 21.1% for all solid media combined.

scholarly journals Comparison of hypertonic saline-sodium hydroxide method with modified Petroff’s method for the decontamination and concentration of sputum samples

2013 ◽  
Vol 2 (3) ◽  
pp. 78-81 ◽  
Author(s):  
SK Chaudhary ◽  
B Mishra
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sodium Hydroxide ◽  
Hypertonic Saline ◽  
P Value ◽  
Chi Square ◽  
Major Health ◽  
Medical College ◽  
Lowenstein Jensen ◽  
Novel Method ◽  
The Difference

INTRODUCTION: Tuberculosis is one of the major health problems particularly in developing countries. For definitive diagnosis of pulmonary tuberculosis identification of tubercle bacilli in sputum by microscopy and culture is essential. For decontamination and concentration of sputum, the commonly used method in the laboratory is Modified Petroff’s method but the Hypertonic saline–sodium hydroxide (HS-SH) method is known to be better for detection of Mycobacterium tuberculosis by culture. This study was aimed to compare a novel method for the improvement of decontamination and concentration of sputum samples. MATERIALS AND METHODS: A total of 50 confirmed smear positive sputum samples from pulmonary TB patients who visited at St. John’s Medical College and Hospital during 2009 to 2010, were processed for the decontamination process. Each sample was decontaminated by Modified Petroff’s and HS-SH method separately. Treated samples were cultured in Lowenstein-Jensen media in microbiology laboratory. RESULTS: The culture positive percents of Mycobacterium tuberculosis in the L-J medium treated with HS-SH and Modified Petroff’s method were 84.0% and 70.0%, respectively. A notable feature is that by HS-SH method more samples were positive by 4th week, statistically significant (Chi- square value-11.26 with p-value < 0.05) compare to Modified Petroff’s method. The difference for 3+ grades of L-J growths found slightly higher by Modified Petroff’s method but at lower grades of growths HS-SH method performed better. CONCLUSIONS: HS-SH method is better for the detection of Mycobacterium tuberculosis by culture when compared with the Modified Petroff’s method. DOI: http://dx.doi.org/10.3126/ijim.v2i3.8664   Int J Infect Microbiol 2013;2(3):78-81

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