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Rapid and Biosecure Diagnostic Test for Tuberculosis

Cell Biochemistry and Biophysics volume 65, pages 173–179 (2013)Cite this article

Abstract

Early and rapid detection of the causative organism is necessary in tuberculosis. We present here an integrated and dedicated molecular biology system for tuberculosis diagnosis. One hundred and eighty-nine (189) biologic specimens from patients strongly suspected by clinical parameters of tuberculosis were studied by Ziehl–Neelsen staining, cultivation on a solid medium, and by a balanced heminested fluorometric PCR system (Orange G3TB) that preserves worker safety and produces a rather pure material free of potential inhibitors. DNA amplification was carried out in a low cost using a tuberculosis thermocycler-fluorometer. The double stranded DNA produced is fluorometrically detected. The whole reaction is carried out in one single tube which is never opened after adding the processed sample, thus minimizing the risk of cross contamination with amplicons. The assay is able to detect 30 bacilli/ml of sample having a 99.8 % inter-assay coefficient of variation. PCR was positive in 36 (18.9 %) tested samples (33 of them were smear-negative). In our study, it yields a preliminary overall sensitivity of 97.4 %. In addition, its overall specificity is 98.7 %. The total run time of the test is 4 h with two and a half real working hours. All PCR-positive samples also had a positive result by microbiological culture and clinical criteria. The results obtained showed that it could be a very useful tool to increase efficiency in detecting the tuberculosis disease in low bacillus inoculum samples. Furthermore, its low cost and friendly usage make it feasible to be used in regions with poor development.

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Authors and Affiliations

  1. Laboratory of Molecular Biology and Pathology, School of Medicine, Buenos Aires University, Buenos Aires, Argentina

    Juan Garberi & Jorge Labrador

  2. Orange Solutions, School of Medicine, Buenos Aires University, Buenos Aires, Argentina

    Federico Garberi, Juan Ezequiel Garberi, Julián Peneipil, Miguel Garberi & Luis Scigliano

  3. Department of Microbiology and Infectious Diseases, School of Medicine, Buenos Aires University, Buenos Aires, Argentina

    Alcides Troncoso

Authors
  1. Juan Garberi
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  2. Jorge Labrador
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  3. Federico Garberi
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  4. Juan Ezequiel Garberi
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  5. Julián Peneipil
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  6. Miguel Garberi
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  7. Luis Scigliano
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  8. Alcides Troncoso
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Correspondence to Juan Garberi.

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Garberi, J., Labrador, J., Garberi, F. et al. Rapid and Biosecure Diagnostic Test for Tuberculosis. Cell Biochem Biophys 65, 173–179 (2013). https://doi.org/10.1007/s12013-012-9413-7

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  • DOI: https://doi.org/10.1007/s12013-012-9413-7

Keywords

  • Tuberculosis
  • Sputum
  • Cerebral spinal fluid
  • Pleural liquid
  • Molecular diagnosis
  • Low cost
  • Real-time PCR