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. 2011;6(9):e24789.
doi: 10.1371/journal.pone.0024789. Epub 2011 Sep 22.

Analytical performance of the Roche LightCycler® Mycobacterium Detection Kit for the diagnosis of clinically important mycobacterial species

Shaheed V Omar  1 , Andreas Roth, Nazir A Ismail, Linda Erasmus, Marthie Ehlers, Marleen Kock, Nuraan Paulse, Halima M Said, Anwar A Hoosen, Udo Reischl
Affiliations

Analytical performance of the Roche LightCycler® Mycobacterium Detection Kit for the diagnosis of clinically important mycobacterial species

Shaheed V Omar et al. PLoS One. 2011.

Abstract

Background: The LightCycler® Mycobacterium Detection Kit based on real-time PCR technology for the detection of Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium kansasii was recently developed. This study evaluated its analytical sensitivity, specificity and reproducibility.

Methodology/principal findings: Plasmid standards were prepared and used to determine the limit of detection. The assay was also performed against organisms other than mycobacteria, other mycobacterial strains and interfering substances to exclude cross-reactivity and interference. Reference standards were prepared and tested to assess the assay's reproducibility. All PCR assays were performed using the LightCycler® 2.0 Instrument. The detection limit for M. tuberculosis was 28 copies per microlitre. Neither cross-reactivity nor interference occurred with non-mycobacterial organisms and substances tested. Overall reproducibility for consecutive measurements, run-to-run, lot-to-lot, day-to-day and laboratory-to-laboratory achieved a coefficient of variance of less than two percent.

Significance: The LightCycler® Mycobacterium Detection kit has shown to be a robust and accurate assay with the potential to be used as a rapid TB diagnostic test.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Probit analysis for determination of the Limit of Detection.
Figure 2
Figure 2. Pooled Melting Temperature (Tm) range of the M. tuberculosis (MycoT), M. avium (MycoA) and M. kansasii (MycoK).
See this image and copyright information in PMC

References

    1. World Health Organization. Global tuberculosis control 2010. 2010. Available: http://www.who.int/tb/publications/global_report/2010/en/index.html. 2011 June 23.
    1. Johnson MM, Waller EA, Leventhal JP. Nontuberculous mycobacterial pulmonary disease. Curr Opin Pulm Med. 2008;14(3):203–210. - PubMed
    1. McGrath EE, McCabe J, Anderson PB American Thoracic SocietyInfectious Diseases Society of America. Guidelines on the diagnosis and treatment of pulmonary non-tuberculous mycobacteria infection. Int J Clin Pract. 2008;62(12):1947–1955. - PubMed
    1. Corbett EL, Blumberg L, Churchyard GJ, Moloi N, Mallory K, et al. Nontuberculous mycobacteria: Defining disease in a prospective cohort of south african miners. Am J Respir Crit Care Med. 1999;160(1):15–21. - PubMed
    1. Andersen P, Doherty TM. The success and failure of BCG - implications for a novel tuberculosis vaccine. Nat Rev Microbiol. 2005;3(8):656–662. - PubMed

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