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Background: Compare the diagnostic sensitivity of ZN staining and CBNAAT using Mycobacterium Culture as a gold standard in BAL fluid among sputum smear negative or non- sputum producing patients with suspected pulmonary tuberculosis.Subjects and Methods:Our prospective observational study was conducted on 76 patients either attending Outpatient Department or being admitted in the department of Pulmonary Medicine, Teerthanker Mahaveer Medical College & Research Centre, Moradabad (UP). BAL sample obtained using a flexible fibre-optic bronchoscope from sputum smear negative or non- sputum producing patients with suspected pulmonary tuberculosis. Sample was divided into three parts and send the BAL fluid for CBNAAT, ZN staining, and Mycobacterium liquid culture and compare the diagnostic sensitivity of ZN staining and CBNAAT using Mycobacterium Culture as a gold standard in BAL fluid.Results: Our study was carried out on 76 patients, ZN staining detected positive and negative in BAL samples among 17.1% and 82.9% respectively. CBNAAT detected positive and negative in BAL samples among 68.72% and 31.6% respectively. Among the positive detected specimens, rifampicin resistance and sensitive were found among 4 (5.56%) and 48 (63.16%) specimens respectively.MTB detected positive and negative in BAL samples among 51.3% and 48.7% of the subjects respectively. The gold standard BAL-MTB liquid culture was used to test the efficacy of ZN staining to detect the BAL specimen. Sensitivity, specificity, PPV, NPV, and Accuracy of ZN in the detection of BAL specimen were 23.08%, 89.19%, 69.23%, 52.38%, and 55.26% respectively. Sensitivity, specificity, PPV, NPV and Accuracy of CBNAAT in detection of BAL specimen were 92.31%, 66.76%, 79.23%, 87.50% and 75% respectively.Conclusion: Gene Xpert MTB/RIF assay is efficient and reliable technique for the smear negative cases. Its simplicity, sensitivity, speed and automation, make this technique a very attractive tool for diagnosis of pulmonary tuberculosis in smear negative cases of TB suspects. Meanwhile it has an added advantage of detection of multi-drug resistant cases.

Tuberculosis-related pleural effusion (TPE) is reported in 12 to 38% of thoracic tuberculosis (TB) cases in the pediatric population. In TPE, the pleural fluid bacilli load is very low, generally resulting in negative acid-fast bacill (AFB) staining and Mycobacterium culture. In the pleural fluid, AFB stain positivity is reported in

Background: The diagnosis of tuberculosis (TB) has mostly been relied on a long-used method called sputum smear microscopy. In 2010, Xpert MTB/RIF assay was approved by the World Health Organization for simultaneous TB diagnosis and detection of resistance. Our current study was undertaken to compare the diagnostic performance of Xpert MTB/RIF assay to auramine staining-based light-emitting diode-Fluorescence Microscopy (LED-FM) considering culture as the gold standard method for pulmonary and extrapulmonary TB. Method: Pulmonary and extrapulmonary specimens of suspected TB patients were examined in this study. From January 2016 to June 2019, sputum, urine, superficial swabs, gastric aspirates, and pleural infusion specimens were collected from new and previously treated TB individuals. Specimens were examined using Xpert MTB/RIF, LED-FM, and Mycobacterium culture techniques to evaluate their performance. Results: A total of 697 suspected TB samples were included in this analysis, and of these, 469 (67.29%) were positive for all three used methods. The overall sensitivities, specificities, and positive and negative predictive values were 99.6%, 62.0%, 88.4%, and 98.2% for Xpert MTB/RIF and 88.0%, 95.6%, 99.0%, and 60.7% for LED-FM, respectively, compared to culture method. Conclusion: The sensitivity of Xpert MTB/RIF assay was observed to be higher than the LED-FM method, thus suggesting this molecular technique as a promising tool for the diagnosis of pulmonary and extrapulmonary TB, which will help in the management of TB infections in developing countries such as Mali.

Here, we report a case of 24-year-old female who experienced fever, weight loss, and headache by the termination of her pregnancy after 24 weeks. She was admitted due to multiple generalized tonic clonic (GTC) seizures and loss of consciousness. Her lab data and histological findings were insignificant as well as negative mycobacterium culture. Her brain MRI detected multiple enhanced brain lesions on T2 weighted images. Overall, she went under empiric anti-TB treatment for a year and had a complete recovery. To our knowledge, this is the first reported case of brain tuberculoma with negative histological findings and brain tissue culture who responded to an empiric anti-TB regimen.

The diagnosis of pulmonary tuberculosis (PTB) is based on a stepwise approach consisting of sputum microscopy and molecular testing (GeneXpert), with the use of Mycobacterium culture in select cases. We analysed a bundled approach of PTB testing, consisting of all three of these tests during the patient's first visit, for all patients fitting into the clinical criteria of PTB. We retrospectively analysed the medical records of all patients who underwent the TB diagnosis bundle during a period of 29 months. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of sputum microscopy and GeneXpert were compared against the gold standard of Mycobacterium culture. The incremental yield of diagnosing PTB by addition of Mycobacterium culture in the diagnostic bundle during the initial testing was also calculated. The bundled approach conferred an advantage in terms of faster clinical decision and increased diagnosis rates, by virtue of the speed of smear microscopy and GeneXpert, combined with the higher sensitivity of culture. The bundle also had the additional benefit of detecting non-tuberculous Mycobacterium (NTM) by the culture method. Hence we feel that the bundled approach ensures fewer number of hospital visits and reduces the potential delays in a clinical decision making.

Background The incidence of Mycobacterium abscessus infections has increased in recent years. Some of these infections are caused by invasive cosmetic procedures. Aims Raising the awareness of cosmetic procedure related Mycobacterium abscessus infection for clinicians. Patients/methods We presented a 28-year-old woman who developed multiple erythema and painful nodules in her lower extremities after injections of botulinum toxin. Results Mycobacterium culture and strain identification of the tissue confirmed Mycobacterium abscessus. Combination antibiotics therapy was given and the lesion healed with scar and pigmentation. Conclusion Mycobacterium abscessus infections following injection of botulinum toxin are rare and easily misdiagnosed as common suppurative infections. Early microbiologic tests are necessary for diagnose. Standardized operation should be performed to avoid this particular infection.

The cultivation of mycobacteria often requires the use of several antibiotics to limit the growth of other rapidly growing micro-flora present in the growth medium. This antibiotic cocktail is one of the most expensive reagents required for mycobacterium culture. Here we present a customized antibiotics mix that is easy to prepare at a fraction of the cost of the commercially available antibiotic mixture that protects against transient flora, which are normally present in lungs, without affecting mycobacterial colony number.

This is a case of a 2-month-old infant who had suppurative lymphadenitis at left axilla after Bacille Calmette Guérin (BCG) vaccination. She presented with a non-healing BCG wound and left axillary swelling associated with one-week fever at 2 months of age. Aspiration under general anaesthesia was done and pus sent for mycobacterium culture grew Mycobacterium tuberculosis complex. The patient improved without anti-tuberculosis treatment.Bangladesh Journal of Medical Science Vol.15(2) 2016 p.283-285

A total of 8,058 male and female mixed-breed goats and 1-4 years of age were slaughtered over a period of 7 months at the public slaughterhouse of Patos city, Paraíba state, in the Northeast region of Brazil; 822 animals were inspected for gross lesions of tuberculosis, and 12 (1.46%) had lesions suggestive of tuberculosis in the mammary gland, lungs, liver and mediastinal, mesenteric, submandibular, parotid and prescapular lymph nodes. Presence of granulomatous lesions was confirmed in the submandibular lymph node of one (8.3%) goat at the histopathological examination and at the mycobacterium culture the same sample was confirmed positive. Isolate was confirmed as belonging to the M. tuberculosis complex by PCR restriction enzyme analysis (PRA). Spoligotyping identified the isolate into spoligotype SB0295 on the M. bovis Spoligotype Database website (www.mbovis.org), and it was classified as M. bovis. The occurrence of M. bovis in goats in this study suggests that this species may be a potential source of infection for humans and should be regarded as a possible problem in the advancement of control and eradication program for bovine tuberculosis in Brazil.

An easy-to-handle Mycobacterium -specific PCR assay for detection of the presence of a wide range of mycobacterial species in clinical samples was evaluated. The performance of the genus probe was compared with the performance of probes specific for Mycobacterium tuberculosis and Mycobacterium avium and with that of standard culture. In addition, the utility of an internal control in monitoring amplification inhibitors was studied. Of 545 respiratory and 325 nonrespiratory specimens (a total of 870 specimens), 58 (6.7%) showed the presence of amplification inhibitors, as determined by a negative result for the internal control. Of these 58 specimens, 31 (53%) were stool specimens; other material, even citrate blood after lysis of erythrocytes, did not pose a problem with regard to inhibition of PCR amplification. Eighty-one of the remaining 812 specimens had a positive Mycobacterium culture result. Of these culture-positive specimens, 58 (71.6%) showed a positive result with the Mycobacterium genus-specific probe. Seventy-two samples had a positive result with the Mycobacterium -specific probe but a negative culture result. Of these 72 samples, 26 samples were regarded as true positive, either because the M. tuberculosis - or M. avium -specific probe was also positive at the same time or because other specimens from the same patient taken at the same time were culture positive. The sensitivity of the Mycobacterium -specific probe was 78.5% and the specificity was 93.5%. This study showed that pretesting of clinical specimens for mycobacteria to the genus level with a Mycobacterium -specific probe offers the routine clinical laboratory the possibility of detecting tuberculous and nontuberculous mycobacteria with one test. Furthermore, specimens testing positive with the genus-specific probe can be immediately identified with species-specific probes.

There are some submerged reefs of rocks in EPTB, which camouflages the problem of it. The first rock is difficulty of diagnostic. EPTB has no any special clinical picture excluding mycobacterium culture – but it is rare in EPTB. The example in figures – 70 % men, died from TB, had tuberculosis of prostate, mostly overlooked alive. For Russia that means about 10,000 men.

Bovine tuberculosis (TB) is a contagious neglected zoonosis of cattle that is prevalent but under-investigated in Cameroon, hence this study was designed to assess the epidemiology of bovine TB in cattle, risks for M. bovis infection in cattle and humans; and public health implications of zoonotic bovine TB in the highlands of Cameroon. A retrospective study of meat inspection records (1994 – 2010) was done to estimate the prevalence of TB lesions in slaughtered cattle in the North West region. The prevalence of bovine TB and anti-bovine TB antibodies in live cattle based on tuberculin skin tests (2 surveys) and immune-chromatographic assays respectively were carried out in the Western and Adamawa highlands of Cameroon. The performance of the tuberculin tests for bovine TB diagnosis in cattle using various tuberculin skin test cut-off points against the detection of anti-bovine TB antibodies (hypothesised risks of exposure) was compared. Suspected TB lesions from slaughtered cattle and infected human sputa were cultured on Lowentein – Jesen and Middlebrook 7H9 media to isolate mycobacteria agents for molecular genotyping using genomic deletion analysis and spoligotyping. Risk factors for exposure and transmission of zoonotic bovine TB infection of cattle and cattle professionals, and its public health significance were determined using structured questionnaires. Seventeen years of meat inspection record revealed that suspect TB lesions were identified in 599 of 129,165 slaughtered cattle at the Bamenda abattoir. The lungs and associated lymph nodes (over 60%) were the most affected tissues. Other results showed that the prevalence of anti-bovine TB antibodies in cattle in the study regions was 37.17%. Chi square statistics revealed that irrespective of the tuberculin test cut-off value (P48), strong associations existed between the detection of anti-bovine TB antibodies and disease status. A 95% confidence interval analysis of the comparative cervical tuberculin tests revealed that the prevalence rates were 4.67% – 7.15%, 12.02% – 15.67% and 20.56% – 24.98% at the ≥ 4mm, ≥ 3mm and ≥ 2mm cut-off points, respectively. Overall, the best test performance was realised at ≥ 3-mm, though the ≥ 2-mm cut-off point predicted more positive reactors. Age, sex, breed and husbandry practices served as significant (P

The mortality rate of patients with miliary tuberculosis is almost 20%. The sensitivity of acid-fast staining method does not exceed 50% for diagnosis of miliary tuberculosis. Nevertheless, the time required for mycobacterium culture results may lead to signiWcant delay in diagnostic procedure. The tuberculin skin test is negative in more than 50% of patients with miliary tuberculosis. This is more distinct in patients who received steroid treatment. The sensitivity (86–89%) and speciWcity (94–98%) of QuantiFERON TB2G (QFT-TB) test is high for the diagnosis of tuberculosis infection and the test is introduced to the clinical applica

Fregnan , G. B. (University of Wisconsin, Madison), D. W. Smith, and H. M. Randall . Biological and chemical studies on mycobacteria. Relationship of colony morphology to mycoside content for Mycobacterium kansasii and Mycobacterium fortuitum . J. Bacteriol. 82: 517–527. 1961.—Using a suitable technique and an adequate medium it was possible to show a unique and uniform type of colony characteristic for Mycobacterium kansasii (photochromogen) and for Mycobacterium fortuitum strains freshly isolated either from patients, or from the soil, or kept in our stock culture collection for several years. New symbols have been proposed to represent these colony types. It was demonstrated that colony morphology is closely related to the specific mycoside present in a given strain; for example, M. kansasii strains showed in each instance colony type K and mycoside A, and M. fortuitum strains showed colony type F and mycoside F. Attention is called to the importance of the technique and the medium used. No change in colony morphology resulted from incubation in the presence of air containing 5 to 10% CO 2 , although this improved growth. Better growth of mycobacteria occurred in the presence of glycerol, or lipids of a human strain of mycobacteria, or sodium bicarbonate, but the specificity of colony form was lost.

Die Erfahrungen mit dem flussigen Nahrsubstrat vonSula werden bei der primaren Zuchtung von Tuberkelbakterien aus sterilem oder verunreinigtem Untersuchungsmaterial als gunstig angesehen. Besonders bei Larynx- oder Fistelabstrichen wird die Flussigkeitskultur als “Vorkultur” bevorzugt. Ebenso vorteilhaft hat sich die Praparation der Abstriche mit einer Penicillinlosung an Stelle der Schwefelsaure erwiesen, ohne dass hierbei eine Schadigung des Nahrsubstrates, oder eine verminderte Ausbeute positiver Kulturen beobachtet worden ware. Die Qualitat des Nahrsubstrates ist zum Teil vom verwendeten Rinderserum abhangig, lasst sich aber durch eine 3-wochige “Reifung” im Kuhlschrank erheblich verbessern. Die bisher gemachten Erfahrungen beruhen auf einer 3-jahrigen Beobachtung.

1. Im Hinblick auf ihre Verwendung als zweite Kohlenstoffquelle neben Glycerin wird der Einflus verschiedener Substrate auf die Ruhe- und Vermehrungsatmung INH-sensibler und-resistenter Bakterien untersucht. 2. Substrate mit einem QO2 zwischen-8,2 und-4 bei ruhenden Bakterien zeigen auch bei sich vermehrenden Keimen eine fur Vermehrungsvorgange charakteristische Sauerstoffaufnahme. Beim Zusatz der anderen Substrate last sich dagegen eine Vermehrung nicht mehr erkennen. 3. Wahrend sich bei der Ruheatmung zwischen INH-sensiblen und-resistenten Bakterien kaum Unterschiede feststellen lassen, ist bei der resistenten Variante die Vermehrungsatmung bei allen gepruften Substraten verzogert. 4. Mit Ausnahme der Ameisensaure entspricht der RQ bei allen Substraten einer vollstandigen Oxydation zu Kohlendioxyd und Wasser. 5. Der RQ fur Brenztrauben- und Oxalessigsaure zeigt zwischen der INH-sensiblen und-resistenten Population statistisch gesicherte Differenzen. Dieser Befund ist mit einer erhohten Decarboxylaseaktivitat der sensiblen Variante zu erklaren.

Fregnan , G. B. (University of Wisconsin, Madison) and D. W. Smith . Description of various colony forms of mycobacteria. J. Bacteriol. 83: 819–827. 1962.—The colony morphology of various mycobacteria, including mammalian and avian strains, Mycobacterium fortuitum, M. kansasii , and representatives of the unclassified groups, is described. A relationship is seen between some colony forms and species of mycobacteria. The reasons for the adoption of a new terminology, based on the words smooth and rough, are discussed.

Seventy-two strains of nonpathogenic acid-fast bacilli isolated from various sources were studied inan attempt to establish any criterion for the classification of these bacilli.On the ground of heat tolerance, urease production, and growth on sorbit-NH4Cl medium, these strains seemed to be classified into four types. Other characters, such as morphology, decomposition of sugar, sensitivity to antibiotics, and allergenic capacity, were of little value for this purpose.The four types were divided according to the following plan.Type I: Survives at 60°C for one hour. Grows at 47°C. Distributed primarily in natural environments. Correspond to Mycobacterium phlei described in Bergey's Manual, 7th edition.Type II: Fails to survive at 60°C for one hour. Grows at 47°C. Most of the strains utilize sodium oxalate. Distributed primarily in natural environments. Corresponds to Mycobacterium smegmatis in Bergey's Manual, 7th edition.Type III: Fails to survive at 60°C for one hour. Fails to grow at 47°C. Grow on sorbit-NH4Cl, medium. Produces urease. Distributed primarily in the natural environments.Type IV: Fails to survive at 60°C for one hour. Fails to grow at 47°C. Fails either to grow on. sorbit-NH4Cl medium or to produce urease. Distributed among animals. Divided into two subtypes.Subtype 1: Fails to grow on sorbit-NH4Cl medium and to produce urease.Subtype 2: Either grows on this medium or produces urease.Fifteen strains of subtype 1 resemble Mycobacterium avium in all biological characters, except pathogenicity.

M. lepraemurium causes rat leprosy, which possesses many of the characteristics of the human disease. So far, attempts to culture the causative organism of either disease in bacteriological media or in tissue cultures have not provided convincing evidence of multiplication.

SUMMARY: Mycobacterium balnei grows best at temperatures about Growth°. 32 occurs at 37° only on certain media. The organism is more susceptible at 37° than at 32° to the inhibitory effects of oleic acid added to a simple defined medium. This appears to explain the failure of more complex media to support growth at the higher temperature.

SUMMARY: Mycobacteria were grown in rocked ⊥ tubes (Monod) in a medium which contained asparagine, glycerol, 1·0% (w/v) Tween 80 and no albumin fraction V. This permitted abundant well-dispersed diffuse growth. The amount of growth could be measured photometrically.

THE work reported here followed a chance observation made while attempting, without success, to grow Mycobacterium leprae in a culture of Entamoeba terrapinae.