Arch Pathol Lab Med—Vol 127, December 2003 Bioelectronic Mutation Detection in Cell Lines—Bernacki et al 1565 Original Articles Bioelectronic Sensor Technology for Detection of Cystic Fibrosis and Hereditary Hemochromatosis Mutations Susan H. Bernacki, PhD; Daniel H. Farkas, PhD; Wenmei Shi, PhD; Vivian Chan, BS; Yenbou Liu, MS; Jeanne C. Beck, PhD; Karen Snow Bailey, PhD; Victoria M. Pratt, PhD; Kristin G. Monaghan, PhD; Karla J. Matteson, PhD; FrederickV. Schaefer, PhD; Michael Friez, PhD; Antony E. Shrimpton, PhD; TimothyT. Stenzel, MD, PhD ● Context.—Bioelectronic sensors, which combine micro- chip and biological components, are an emerging technol- ogy in clinical diagnostic testing. An electronic detection platform using DNA biochip technology (eSensor) is under development for molecular diagnostic applications. Owing to the novelty of these devices, demonstrations of their successful use in practical diagnostic applications are lim- ited. Objective.—To assess the performance of the eSensor bioelectronic method in the validation of 6 Epstein-Barr virus–transformed blood lymphocyte cell lines with clini- cally important mutations for use as sources of genetic ma- terial for positive controls in clinical molecular genetic testing. Two cell lines carry mutations in the CFTR gene (cystic fibrosis), and 4 carry mutations in the HFE gene (hereditary hemochromatosis). Design.—Samples from each cell line were sent for ge- notype determination to 6 different molecular genetic test- ing facilities, including the laboratory developing the DNA biochips. In addition to the bioelectronic method, at least 3 different molecular diagnostic methods were used in the analysis of each cell line. Detailed data were collected from the DNA biochip output, and the genetic results were compared with those obtained using the more established methods. Results.—We report the successful use of 2 applications of the bioelectronic platform, one for detection of CFTR mutations and the other for detection of HFE mutations. In all cases, the results obtained with the DNA biochip were in concordance with those reported for the other methods. Electronic signal output from the DNA biochips clearly differentiated between mutated and wild-type al- leles. This is the first report of the use of the cystic fibrosis detection platform. Conclusions.—Bioelectronic sensors for the detection of disease-causing mutations performed well when used in a ‘‘real-life’’ situation, in this case, a validation study of pos- itive control blood lymphocyte cell lines with mutations of public health importance. This study illustrates the practi- cal potential of emerging bioelectronic DNA detection technologies for use in current molecular diagnostic ap- plications. (Arch Pathol Lab Med. 2003;127:1565–1572) E merging technologies that incorporate biological ele- ments into microchip-based devices have resulted in the development of novel clinical diagnostic methods, par- Accepted for publication July 29, 2003. From the Department of Pathology, Duke University MedicalCenter, Durham, NC (Drs Bernacki and Stenzel); Motorola Life Sciences, Pas- adena, Calif (Drs Farkas and Shi, Ms Chan, and Mr Liu); Coriell Institute for Medical Research, Camden, NJ (Dr Beck); the Department of Lab- oratory Medicine, Mayo Clinic, Rochester, Minn (Dr Snow Bailey); Lab- oratory Corporation of America, Research Triangle Park, NC (Dr Pratt); the Department of Medical Genetics, Henry Ford Hospital, Detroit, Mich (Dr Monaghan); the Departments of Medical Genetics and Pa- thology, University ofTennessee Medical Center, Knoxville (Dr Matte- son); Chapman Institute of Medical Genetics, Tulsa, Okla (Dr Schaefer); Greenwood Genetic Center, Greenwood, SC (Dr Friez); and the De- partment of Clinical Pathology, SUNY Upstate Medical University,Syr- acuse, NY (Dr Shrimpton). Dr Farkas is now with the Department of Pathology, Baylor College of Medicine, Houston,Tex; Ms Chan is now with the Molecular Pathology Department, Stanford Hospital and Clin- ics, Stanford, Calif; Dr Snow Bailey is now with the Department of Diagnostic Genetics, Auckland Hospital, Auckland, New Zealand; Dr Pratt is now with Nichols Institute, Quest Diagnostics, Chantilly,Va; Dr Stenzel is now with Vysis, Inc, an Abbott Laboratories Company, Downers Grove, Ill; and Motorola Life Sciences is now Clinical MicroSensors, a Motorola Company. ticularly in the area of clinical molecular genetics (re- viewed in McGlennen 1 ). Nucleic acid–based bioelectronic sensors are one of many applications of these technologies that hold promise for low-cost, automated diagnostic sys- tems. Nucleic acid–based bioelectronic detection involves the generation of an electronic signal mediated by nucleic acid hybridization and serves as the basis for the eSensor DNA detection technology (Motorola Life Sciences, Pasadena, Calif). 2–4 The technical specifications of the eSensor plat- form have been described previously. 4 The method is based on hybridization between 3 single-stranded DNA molecules: a DNA capture probe, a target DNA, and an electrically active DNA signal probe. DNA capture probes of approximately 21 bases are immobilized in specified patterns on low-density arrays of gold electrodes on print- The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US government. Reprints: Timothy T. Stenzel, MD, PhD,Vysis, Inc, 3100 Woodcreek Dr, Downers Grove, IL 60515-5400 (e-mail: timothy.stenzel@vysis. com).