JMD TIDES - Oligonucleotide and Peptide - May 18-21, 2008, Las Vegas, NV
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JMD 2003, Vol. 5, No. 4
Copyright © 2003 American Society for Investigative Pathology & Association for Molecular Pathology

Establishment of Stably EBV-Transformed Cell Lines from Residual Clinical Blood Samples for Use in Performance Evaluation and Quality Assurance in Molecular Genetic Testing

Susan H. Bernacki*, Ana K. Stankovic{dagger}, Laurina O. Williams{dagger}, Jeanne C. Beck{ddagger}, James E. Herndon§, Karen Snow-Bailey, Thomas W. Prior||, Karla J. Matteson**, Linda M. Wasserman{dagger}{dagger}, Eugene C. Cole{ddagger}{ddagger} and Timothy T. Stenzel*

From the Molecular Diagnostics Laboratory, * Department of Pathology, and the Comprehensive Cancer Center, § Duke University Medical Center, Durham, North Carolina; the Public Health Practice Program Office, {dagger} Centers for Disease Control and Prevention, Atlanta, Georgia; Coriell Cell Repositories, {ddagger} Coriell Institute for Medical Research, Camden, New Jersey; the Molecular Genetics Laboratory, ¶ Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota; the Department of Pathology, || Ohio State University Hospital, Columbus, Ohio; the Department of Medical Genetics and the Department of Pathology, ** University of Tennessee Medical Center, Knoxville, Tennessee; the Department of Medicine, {dagger}{dagger} University of California San Diego, La Jolla, California; and the Department of Health Science, {ddagger}{ddagger} Brigham Young University, Provo, Utah

Positive control materials for clinical molecular genetic testing applications are currently in critically short supply or non-existent for many genetically based diseases of public health importance. Here we demonstrate that anonymous, residual, clinical blood samples are potential sources of viable lymphocytes for establishing Epstein-Barr virus (EBV)-transformed blood lymphocyte cell lines. We attempted to transform 34 residual blood samples, and analyzed transformation success with respect to sample age, anticoagulant, storage temperature, volume, hemolysis, and patient age and sex. In univariate analysis, sample age was significantly associated with transformation success (P = 0.002). The success rate was 67% (6 of 9) for samples 1 to 7 days old, 38% (3 of 8) for samples 8 to 14 days old and 0% for samples 15 to 21 (0 of 11) days old. When we controlled for sample age in multivariate logistic regression, anticoagulant and storage temperature approached significance (P = 0.070 and 0.087, respectively; samples in acid citrate dextrose (ACD) and refrigerated samples were more likely to transform). Based on these findings, we suggest that samples collected in either ACD or ethylene diamine tetraacetic acid, and up to 14 days old (refrigerated) or 7 days old (stored ambient), are reasonable candidates for EBV transformation. The transformation rate for samples that met these criteria was 63% (10 of 16). Implementation of this process could help alleviate the shortage of positive control materials for clinical molecular genetic testing.




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