Inter-laboratory validation of PCR-based detection of Mycobacterium tuberculosis in formalin-fixed, paraffin-embedded tissues.
The present study is based on the initiative for quality assurance in pathology of the German Society of Pathology and the Professional Association of German Pathologists. Four panel laboratories with experience and expertise in polymerase chain reaction (PCR) detection of Mycobacterium tuberculosis were selected to establish the prerequisites for continuous external laboratory trials, in particular, by providing pre-tested specimens and evaluation criteria for participating institutes. In the first step, the four panel laboratories performed an internal trial to test their own reliability and reproducibility. Paraffin sections and DNA preparations from 34 tissues (25 clinical specimens and 9 controls) totalling to 66 samples were evaluated by each panel institute according to their own protocols. The methodologies differed and are described in detail. Despite these differences, a high degree of inter-laboratory reliability was achieved. In this report, we summarise our results including the correlation with the histology and provide recommendations for applying PCR-based methodology for the detection of mycobacterial DNA in surgical specimens. Supplementary data are available online at http://www.charite.de/ch/patho (rubric "Forschung"). Pre-tested specimens are now available for the external trial and can be ordered from the steering institute via Oligene (http://www.oligene.com/). All molecular pathology laboratories are invited to participate in this quality assurance initiative.
Schewe
Christiane
C
Charité-Campus Mitte, Institute of Pathology, Universitätsmedizin Berlin, Schumannstr. 20-21, 10098 Berlin, Germany. christiane.schewe@charite.de
Goldmann
Torsten
T
Grosser
Marianne
M
Zink
Albert
A
Schlüns
Karsten
K
Pahl
Stefan
S
Ulrichs
Timo
T
Kaufmann
Stefan H E
SH
Nerlich
Andreas
A
Baretton
Gustavo B
GB
Dietel
Manfred
M
Vollmer
Ekkehard
E
Petersen
Iver
I
D000818:Animals; D004269:DNA, Bacterial / Q000032:analysis*; D005557:Formaldehyde; D006801:Humans; D008828:Microbiological Techniques / Q000592:standards; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D016612:Paraffin Embedding; D010338:Pathology, Clinical / Q000592:standards*; D016133:Polymerase Chain Reaction / Q000379:methods; D011785:Quality Assurance, Health Care / Q000592:standards*; D015203:Reproducibility of Results; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis / Q000382:microbiology
D004269:DNA, Bacterial; D005557:Formaldehyde