Detection of Mycobacterium tuberculosis complex with Real Time PCR: comparison of different primer-probe sets based on the IS6110 element.
The sensitivity and specificity to detect Mycobacterium tuberculosis complex of four Real Time PCR primer-probe sets was compared. Three sets targeted nearly the same location on the IS6110 sequence and set 4 targeted a location 200 bp downstream on IS6110. Real Time PCR's with sets 1, 2 and 3 were carried out with co-amplification of a modified target as an internal amplification control. By testing identical DNA samples it was shown that small changes in primer and probe sequences result in differences in the performance of the assays, regarding analytical sensitivity and specificity.
Savelkoul
Paul H M
PH
Department of Medical Microbiology and Infection Control, VU University Medical Center, Amsterdam, The Netherlands. p.savelkoul@vumc.nl
Catsburg
Arnold
A
Mulder
Sije
S
Oostendorp
Ludo
L
Schirm
Jurjen
J
Wilke
Hans
H
van der Zanden
Adri G M
AG
Noordhoek
Gerda T
GT
D017931:DNA Primers / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000382:microbiology*
D017931:DNA Primers; D015342:DNA Probes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial