title: Isothermal multiple displacement amplification: a methodical approach enhancing molecular routine diagnostics of microcarcinomas and small biopsies.
abstract: METHODS
  Isothermal multiple displacement amplification (IMDA) can be a powerful tool in molecular routine diagnostics for homogeneous and sequence-independent whole-genome amplification of notably small tumor samples, eg, microcarcinomas and biopsies containing a small amount of tumor. Currently, this method is not well established in pathology laboratories. We designed a study to confirm the feasibility and convenience of this method for routine diagnostics with formalin-fixed, paraffin-embedded samples prepared by laser-capture microdissection.
  
  
  RESULTS
  A total of 250 μg DNA (concentration 5 μg/μL) was generated by amplification over a period of 8 hours with a material input of approximately 25 cells, approximately equivalent to 175 pg of genomic DNA. In the generated DNA, a representation of all chromosomes could be shown and the presence of elected genes relevant for diagnosis in clinical samples could be proven. Mutational analysis of clinical samples could be performed without any difficulty and showed concordance with earlier diagnostic findings.
  
  
  CONCLUSIONS
  We established the feasibility and convenience of IMDA for routine diagnostics. We also showed that small amounts of DNA, which were not analyzable with current molecular methods, could be sufficient for a wide field of applications in molecular routine diagnostics when they are preamplified with IMDA.
authors.affiliation: Institute of Pathology, University Hospital Essen, Essen, Germany.
authors.forename: Fabian D
authors.initials: FD
authors.lastname: Mairinger
authors.affiliation: Ruhrlandklinik, West German Lung Center, University Hospital Essen, Essen, Germany.
authors.forename: Robert Fh
authors.initials: RF
authors.lastname: Walter
authors.affiliation: Institute of Pathology, University Hospital Cologne, Cologne, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Vollbrecht
authors.affiliation: Institute of Pathology, University Hospital Essen, Essen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hager
authors.affiliation: Institute of Pathology, University Hospital Essen, Essen, Germany.
authors.forename: Karl
authors.initials: K
authors.lastname: Worm
authors.affiliation: Institute of Pathology, University Hospital Essen, Essen, Germany.
authors.forename: Saskia
authors.initials: S
authors.lastname: Ting
authors.affiliation: Institute of Pathology, University Hospital Essen, Essen, Germany.
authors.forename: Jeremias
authors.initials: J
authors.lastname: Wohlschläger
authors.affiliation: Pulmonary Department, Oncology Unit, G Papanikolaou General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece.
authors.forename: Paul
authors.initials: P
authors.lastname: Zarogoulidis
authors.affiliation: Pulmonary Department, Oncology Unit, G Papanikolaou General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Zarogoulidis
authors.affiliation: Institute of Pathology, University Hospital Essen, Essen, Germany.
authors.forename: Kurt W
authors.initials: KW
authors.lastname: Schmid
keywords: biopsies; isothermal; isothermal multiple displacement amplification; microcarcinomas; microdissection; routine diagnostics; whole-genome amplification