title: Analytical performance of the Roche LightCycler® Mycobacterium Detection Kit for the diagnosis of clinically important mycobacterial species.
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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Medical Microbiology, University of Pretoria, Pretoria, South Africa. shaheed.vally@gmail.com
authors.forename: Shaheed V
authors.initials: SV
authors.lastname: Omar
authors.forename: Andreas
authors.initials: A
authors.lastname: Roth
authors.forename: Nazir A
authors.initials: NA
authors.lastname: Ismail
authors.forename: Linda
authors.initials: L
authors.lastname: Erasmus
authors.forename: Marthie
authors.initials: M
authors.lastname: Ehlers
authors.forename: Marleen
authors.initials: M
authors.lastname: Kock
authors.forename: Nuraan
authors.initials: N
authors.lastname: Paulse
authors.forename: Halima M
authors.initials: HM
authors.lastname: Said
authors.forename: Anwar A
authors.initials: AA
authors.lastname: Hoosen
authors.forename: Udo
authors.initials: U
authors.lastname: Reischl
mesh_terms: D001431:Bacteriological Techniques / Q000592:standards; D019411:Clinical Laboratory Techniques / Q000592:standards; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000592:standards; D003937:Diagnosis, Differential; D006801:Humans; D009164:Mycobacterium Infections / Q000175:diagnosis / Q000382:microbiology*; D009162:Mycobacterium avium / Q000235:genetics* / Q000302:isolation & purification; D019909:Mycobacterium kansasii / Q000235:genetics* / Q000302:isolation & purification; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity; D044366:Transition Temperature
chemical_list: D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic