title: Inter-laboratory validation of PCR-based detection of Mycobacterium tuberculosis in formalin-fixed, paraffin-embedded tissues.
abstract: 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.
authors.affiliation: Charité-Campus Mitte, Institute of Pathology, Universitätsmedizin Berlin, Schumannstr. 20-21, 10098 Berlin, Germany. christiane.schewe@charite.de
authors.forename: Christiane
authors.initials: C
authors.lastname: Schewe
authors.forename: Torsten
authors.initials: T
authors.lastname: Goldmann
authors.forename: Marianne
authors.initials: M
authors.lastname: Grosser
authors.forename: Albert
authors.initials: A
authors.lastname: Zink
authors.forename: Karsten
authors.initials: K
authors.lastname: Schlüns
authors.forename: Stefan
authors.initials: S
authors.lastname: Pahl
authors.forename: Timo
authors.initials: T
authors.lastname: Ulrichs
authors.forename: Stefan H E
authors.initials: SH
authors.lastname: Kaufmann
authors.forename: Andreas
authors.initials: A
authors.lastname: Nerlich
authors.forename: Gustavo B
authors.initials: GB
authors.lastname: Baretton
authors.forename: Manfred
authors.initials: M
authors.lastname: Dietel
authors.forename: Ekkehard
authors.initials: E
authors.lastname: Vollmer
authors.forename: Iver
authors.initials: I
authors.lastname: Petersen
mesh_terms: 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
chemical_list: D004269:DNA, Bacterial; D005557:Formaldehyde