Frederick V Schaefer

Publications (4)26.29 Total impact

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  Article: Characterization of publicly available lymphoblastoid cell lines for disease-associated mutations in 11 genes.

  Susan H Bernacki, Jeanne C Beck, Kasinathan Muralidharan, Frederick V Schaefer, Antony E Shrimpton, Kristy L Richie, Barbara C Levin, Genevieve Pont-Kingdon, Timothy T Stenzel
  Clinical Chemistry 12/2005; 51(11):2156-9. · 7.91 Impact Factor
• Article: Genetically characterized positive control cell lines derived from residual clinical blood samples.

  Susan H Bernacki, Jeanne C Beck, Ana K Stankovic, Laurina O Williams, Jean Amos, Karen Snow-Bailey, Daniel H Farkas, Michael J Friez, Feras M Hantash, Karla J Matteson, [......], Elizabeth M Rohlfs, Frederick V Schaefer, Antony E Shrimpton, Elaine B Spector, Catherine A Stolle, Charles M Strom, Stephen N Thibodeau, Eugene C Cole, Barbara K Goodman, Timothy T Stenzel
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  ABSTRACT: Positive control materials for clinical diagnostic molecular genetic testing are in critically short supply. High-quality DNA that closely resembles DNA isolated from patient specimens can be obtained from Epstein-Barr virus (EBV)-transformed peripheral blood lymphocyte cell lines. Here we report the development of a process to (a) recover residual blood samples with clinically important mutations detected during routine medical care, (b) select samples likely to provide viable lymphocytes for EBV transformation, (c) establish stable cell lines and confirm the reported mutation(s), and (d) validate the cell lines for use as positive controls in clinical molecular genetic testing applications. A network of 32 genetic testing laboratories was established to obtain anonymous, residual clinical samples for transformation and to validate resulting cell lines for use as positive controls. Three panel meetings with experts in molecular genetic testing were held to evaluate results and formulate a process that could function in the context of current common practices in molecular diagnostic testing. Thirteen laboratories submitted a total of 113 residual clinical blood samples with mutations for 14 genetic disorders. Forty-one EBV-transformed cell lines were established. Thirty-five individual point and deletion mutations were shown to be stable after 20 population doublings in culture. Thirty-three cell lines were characterized for specific mutations and validated for use as positive controls in clinical diagnostic applications. A process for producing and validating positive control cell lines from residual clinical blood samples has been developed. Sustainable implementation of the process could help alleviate the current shortage of positive control materials.
  Clinical Chemistry 12/2005; 51(11):2013-24. · 7.91 Impact Factor
• Article: Multicenter characterization and validation of the intron-8 poly(T) tract (IVS8-T) status in 25 Coriell cell repository cystic fibrosis reference cell lines for cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation assays.

  Siby Sebastian, Silvia G Spitzer, Leonard E Grosso, Jean Amos, Frederick V Schaefer, Elaine Lyon, Daynna J Wolff, Atieh Hajianpour, Annette K Taylor, Alison Millson, Timothy T Stenzel
  Clinical Chemistry 02/2004; 50(1):251-4. · 7.91 Impact Factor
• Article: Bioelectronic sensor technology for detection of cystic fibrosis and hereditary hemochromatosis mutations.

  Susan H Bernacki, Daniel H Farkas, Wenmei Shi, Vivian Chan, Yenbou Liu, Jeanne C Beck, Karen Snow Bailey, Victoria M Pratt, Kristin G Monaghan, Karla J Matteson, Frederick V Schaefer, Michael Friez, Antony E Shrimpton, Timothy T Stenzel
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  ABSTRACT: Bioelectronic sensors, which combine microchip and biological components, are an emerging technology 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 limited. To assess the performance of the eSensor bioelectronic method in the validation of 6 Epstein-Barr virus-transformed blood lymphocyte cell lines with clinically important mutations for use as sources of genetic material 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). Samples from each cell line were sent for genotype determination to 6 different molecular genetic testing 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. 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 alleles. This is the first report of the use of the cystic fibrosis detection platform. 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 positive control blood lymphocyte cell lines with mutations of public health importance. This study illustrates the practical potential of emerging bioelectronic DNA detection technologies for use in current molecular diagnostic applications.
  Archives of pathology & laboratory medicine 01/2004; 127(12):1565-72. · 2.58 Impact Factor