title: Automation of polymerase chain reaction tests. Reduction of human errors leading to contamination.
abstract: We compared the performance over 21 months of manually performed polymerase chain reaction (PCR)-based DNA analysis experiments with 25 months of automated PCR performed by a Zymark robotic system. Automation of the PCR technique resulted in a sixfold reduction in the number of experiments reporting carryover contamination and decreased the overall rate of contamination among total reactions 68-fold. Whereas contamination occurrences among manual experiments were evenly dispersed over the study interval and correlated with the lack of experience of laboratory personnel, the contamination that occurred with the robotic system was confined to the first 10 months of operation. In manual experiments, many of the 81 no-target false positives were sufficiently strong to result in the invalidation of 151 samples and positive controls. The seven no-target control false positives in the automated system were weak bands that were easily subtracted as background. Because none of the negative samples had DNA bands, no sample on the automated system has ever been invalidated as a result of contamination. Automation of PCR tests appears to offer great promise in reducing contamination to acceptable levels (e.g., < or = 0.1%).
authors.affiliation: Department of Pathology, University of Iowa College of Medicine, Iowa City 52242, USA.
authors.forename: W W
authors.initials: WW
authors.lastname: Wilke
authors.forename: R N
authors.initials: RN
authors.lastname: Jones
authors.forename: L D
authors.initials: LD
authors.lastname: Sutton
mesh_terms: D004247:DNA / Q000032:analysis*; D004866:Equipment Contamination; D006801:Humans; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D015203:Reproducibility of Results; D012371:Robotics; D012680:Sensitivity and Specificity
chemical_list: D004247:DNA