Cobas taqman mtb test

Manufactured by Roche

Sourced in Switzerland

The Cobas TaqMan MTB test is a qualitative in vitro diagnostic test for the direct detection of Mycobacterium tuberculosis complex DNA from clinical specimens. The test utilizes real-time PCR technology to amplify and detect specific DNA sequences, providing results that aid in the diagnosis of tuberculosis.

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Lab products found in correlation

Mgit 960 system, by BD (2 mentions) Cobas mtb test, by Roche (2 mentions) Mycobacterial growth indicator tube, by BD (1 mentions) Genexpert mtb rif, by Cepheid (1 mentions)

5 protocols using cobas taqman mtb test

Diagnosing Pediatric Tuberculosis

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Clinical and anthropometric data were recorded. A chest radiograph (CXR), anterioposterior view, was interpreted by 3 independent radiologists. Agreement by 2 radiologists was required to classify a CXR as consistent with TB. A TST was performed by a trained nurse/doctor (2 TU/0.1 mL tuberculin; Span Diagnostics Ltd, Gujarat, India) and read after 48 hours. Peripheral blood (3 mL) was drawn for the QFT (Cellestis/Qiagen, Carnegie, Victoria, Australia) which was performed according to the manufacturer’s instructions.
Gastric aspirates and induced sputa were collected on 2 consecutive days for fluorescent microscopy (Auramine) and culture on solid (Löwenstein-Jensen) and liquid (Mycobacterial Growth Indicator Tube, BD) medium. Positive results were confirmed by Ziehl-Neelsen staining and speciated using the HAIN kit (GenoType MTBC, Ver1, Hain Life Sciences, Germany). Direct PCR (The COBAS TaqMan MTB Test, Roche 2007) was done on all culture-negative specimens in children with CXR consistent with TB. The HIV status was not assessed, but data from a household contact study conducted in the same study area during the period 2010–2012 found a HIV prevalence <1% (TB Trials Study Group, unpublished data).

Jenum S., Selvam S., Mahelai D., Jesuraj N., Cárdenas V., Kenneth J., Hesseling A.C., Doherty T.M., Vaz M, & Grewal H.M. (2014). Influence of Age and Nutritional Status on the Performance of the Tuberculin Skin Test and QuantiFERON-TB Gold In-Tube in Young Children Evaluated for Tuberculosis in Southern India. The Pediatric Infectious Disease Journal, 33(10), e260-e269.

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Comparative Evaluation of TB Diagnostic Platforms

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All compared studies had to be performed with a common CE-IVD certified, commercially available platform. In addition, specimens had to be obtained in a comparable epidemiological setting: the standard used in this work is a low incidence or notification rate of <10 TB cases /100.000 population^{23 (link)}. For a better comparability of study results only respiratory specimens were taken into account. A PubMed/NCBI research was performed using the search-terms: “evaluation” and “performance” with each of the chosen platforms’ names: GeneXpert MTB/RIF (Cepheid), ProbeTec ET Mycobacterium tuberculosis complex Direct Test (DTB) (Becton- Dickinson), COBAS Taq-Man MTB Test (Roche, Basel, Switzerland) altogether and separately in every combination. Only publications in English were included.

Kohlmorgen B., Elias J, & Schoen C. (2017). Improved performance of the artus Mycobacterium tuberculosis RG PCR kit in a low incidence setting: a retrospective monocentric study. Scientific Reports, 7, 14127.

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Isolating and Culturing Virulent M. tuberculosis

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M. tuberculosis were obtained from a clinical virulent strain identified by the Mycobacteriology Laboratory, MacKay Memorial Hospital. Firstly, M. tuberculosis was identified by a MPT64 rapid test, arylsulfatase test, and nitrate reductase assay. The identified strain was further confirmed by a commercial TB-PCR kit (COBAS, Taqman MTB test, Roche). Seed stocks of the collected strain were maintained in small aliquots at -80 °C. The M. tuberculosis used throughout this study was prepared from the seed stocks by culturing on 7H11 agar plate for 3 weeks at 37 °C in 10 % CO2 humidified atmosphere. The culture medium used was a 7H9 broth supplemented with 0.2 % glycerol, 0.05 % Tween-80 and 10 % oleic acid-albumin-dextrose-catalase enrichment (Difco, Becton Dickinson and Company, MD, USA) to an optical density at 600 nm of 0.3 and used as the inoculum.

Kuo C.P., Chang K.S., Hsu J.L., Tsai I.F., Lin A.B., Wei T.Y., Wu C.L, & Lu Y.T. (2016). Analysis of the immune response of human dendritic cells to Mycobacterium tuberculosis by quantitative proteomics. Proteome Science, 14, 5.

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Diagnostic Evaluation of Mycobacterial Infections

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We performed AFB staining with auramine–rhodamine fluorescent stain and then confirmed with Ziehl–Neelsen staining. We graded the staining results according to the American Thoracic Society/Centers for Disease Control and Prevention guidelines.[9 (link)] Then, specimens classified with AFB smear staining results in Grades 1–4 were defined as smear-positive. All clinical specimens were cultured on solid and liquid media for 6 weeks. We inoculated the decontaminated samples into a mycobacterium growth indicator tube (MGIT 960 system; Becton Dickinson, Sparks, MD) and inoculated onto 3% Ogawa agar (Shenyang, Seoul, Korea). All positive cultures were examined by AFB smear to confirm the presence of AFB and to rule out contamination. The Cobas TaqMan MTB test (Cobas MTB test) (Roche Diagnostics, Basel, Switzerland) is a real-time PCR method for analyzing sputum and pleural effusion samples.[9 (link)]

Zhao T., Chen B., Xu Y, & Qu Y. (2020). Clinical and pathological differences between polymorphonuclear-rich and lymphocyte-rich tuberculous pleural effusion. Annals of Thoracic Medicine, 15(2), 76-83.

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Mycobacterial Detection and Identification

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AFB staining was performed with an auramine-rhodamine fluorescent stain, followed by confirmation with Ziehl-Neelsen staining. Staining results were graded according to the American Thoracic Society/Centers for Disease Control and Prevention guidelines [17 (link)]. Specimens in which the AFB smear results were categorized as grades 1 to 4 were defined as smear-positive. All clinical specimens were cultured on both solid and liquid medium for 6 weeks. To this end, decontaminated samples were inoculated into a mycobacterial growth indicator tube (MGIT 960 system; Becton Dickinson, Sparks, MD) and also onto 3% Ogawa agar (Shinyang, Seoul, Korea). All positive cultures were subjected to an AFB smear to confirm the presence of AFB and to exclude contamination. The Cobas TaqMan MTB test (Cobas MTB test) (Roche Diagnostics, Basel, Switzerland), which is a real-time PCR assay, was used to analyze sputum and pleural effusion samples [18 (link)].

Choi H., Chon H.R., Kim K., Kim S., Oh K.J., Jeong S.H., Jung W.J., Shin B., Jhun B.W., Lee H., Park H.Y, & Koh W.J. (2016). Clinical and Laboratory Differences between Lymphocyte- and Neutrophil-Predominant Pleural Tuberculosis. PLoS ONE, 11(10), e0165428.

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