Skip to main content
U.S. flag

An official website of the United States government

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.

Genetically Characterized Positive Control Cell Lines Derived from Residual Clinical Blood Samples



S H. Bernacki, J C. Beck, A K. Stankovic, L O. Williams, J Amos, K Snow-Bailey, D H. Farkas, M J. Friez, F M. Hantash, Karla J. Matteson, M G. Monaghan, Kasinathan Muralidharan, V M. Pratt, T W. Prior, K L. Ritchie, Barbara C. Levin, Elizabeth M. Rohlfs, Fredrick V. Schaefer, A E. Shrimpton, Elaine B. Spector, C A. Stolle, C M. Strom, S N. Thibodeau, E C. Cole, B K. Goodman, T T. Stenzel


Background: 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 provided by Epstein Barr virus (EBV) transformed peripheral blood lymphocyte cell lines. Here we report the development of a process to: 1) recover residual blood samples with clinically important mutations detected during patients routine medical care; 2) select samples likely to provide viable lymphocytes for EBV transformation; 3) establish stable cell lines and confirm the reported mutation(s); 4) validate the cell lines for use as positive controls in clinical molecular genetic testing applications.Method: A network of 32 genetic testing laboratories was established in order 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.Results: Thirteen laboratories submitted a total of 113 residual clinical blood samples with mutations for 14 genetic disorders. 41 EBV transformed cell lines were established. 35 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.Conclusion: 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
No. 11


Cell repository, clinical genetics, cystic fibrosis cystic, fragile X, genetic diseases, hemochromatosis, hemoglobin C, Huntington Disease, sickle cell


Bernacki, S. , Beck, J. , Stankovic, A. , Williams, L. , Amos, J. , Snow-Bailey, K. , Farkas, D. , Friez, M. , Hantash, F. , Matteson, K. , Monaghan, M. , Muralidharan, K. , Pratt, V. , Prior, T. , Ritchie, K. , Levin, B. , Rohlfs, E. , Schaefer, F. , Shrimpton, A. , Spector, E. , Stolle, C. , Strom, C. , Thibodeau, S. , Cole, E. , Goodman, B. and Stenzel, T. (2005), Genetically Characterized Positive Control Cell Lines Derived from Residual Clinical Blood Samples, Clinical Chemistry (Accessed February 22, 2024)
Created October 31, 2005, Updated October 12, 2021