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 Matteson, Kristin G. Monaghan, Kasinathan Muralidharan, Victoria M. Pratt, Thomas W. Prior, Kristy L. Richie, Barbara C. Levin, Elizabeth M. Rohlfs, Frederick V. Schaefer, Antony E. Shrimpton, Elaine B. Spector & 6 others Catherine A. Stolle, Charles M. Strom, Stephen N. Thibodeau, Eugene C. Cole, Barbara K. Goodman, Timothy T. Stenzel

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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 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. Methods: 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. Results: 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. Conclusions: 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.

Original languageEnglish (US)
Pages (from-to)2013-2024
Number of pages12
JournalClinical Chemistry
Volume51
Issue number11
DOIs
StatePublished - Nov 1 2005

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Blood
Cells
Genetic Testing
Cell Line
Human Herpesvirus 4
Molecular Biology
Viruses
Mutation
Testing
Lymphocytes
Molecular Diagnostic Techniques
Transformed Cell Line
Inborn Genetic Diseases
Sequence Deletion
DNA
Point Mutation
Health care
Cell culture
Blood Cells
Population

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Bernacki, S. H., Beck, J. C., Stankovic, A. K., Williams, L. O., Amos, J., Snow-Bailey, K., ... Stenzel, T. T. (2005). Genetically characterized positive control cell lines derived from residual clinical blood samples. Clinical Chemistry, 51(11), 2013-2024. https://doi.org/10.1373/clinchem.2005.048694

Genetically characterized positive control cell lines derived from residual clinical blood samples. / Bernacki, Susan H.; Beck, Jeanne C.; Stankovic, Ana K.; Williams, Laurina O.; Amos, Jean; Snow-Bailey, Karen; Farkas, Daniel H.; Friez, Michael J.; Hantash, Feras M.; Matteson, Karla; Monaghan, Kristin G.; Muralidharan, Kasinathan; Pratt, Victoria M.; Prior, Thomas W.; Richie, Kristy L.; Levin, Barbara C.; Rohlfs, Elizabeth M.; Schaefer, Frederick V.; Shrimpton, Antony E.; Spector, Elaine B.; Stolle, Catherine A.; Strom, Charles M.; Thibodeau, Stephen N.; Cole, Eugene C.; Goodman, Barbara K.; Stenzel, Timothy T.

In: Clinical Chemistry, Vol. 51, No. 11, 01.11.2005, p. 2013-2024.

Research output: Contribution to journalArticle

Bernacki, SH, Beck, JC, Stankovic, AK, Williams, LO, Amos, J, Snow-Bailey, K, Farkas, DH, Friez, MJ, Hantash, FM, Matteson, K, Monaghan, KG, Muralidharan, K, Pratt, VM, Prior, TW, Richie, KL, Levin, BC, Rohlfs, EM, Schaefer, FV, Shrimpton, AE, Spector, EB, Stolle, CA, Strom, CM, Thibodeau, SN, Cole, EC, Goodman, BK & Stenzel, TT 2005, 'Genetically characterized positive control cell lines derived from residual clinical blood samples', Clinical Chemistry, vol. 51, no. 11, pp. 2013-2024. https://doi.org/10.1373/clinchem.2005.048694
Bernacki SH, Beck JC, Stankovic AK, Williams LO, Amos J, Snow-Bailey K et al. Genetically characterized positive control cell lines derived from residual clinical blood samples. Clinical Chemistry. 2005 Nov 1;51(11):2013-2024. https://doi.org/10.1373/clinchem.2005.048694
Bernacki, Susan H. ; Beck, Jeanne C. ; Stankovic, Ana K. ; Williams, Laurina O. ; Amos, Jean ; Snow-Bailey, Karen ; Farkas, Daniel H. ; Friez, Michael J. ; Hantash, Feras M. ; Matteson, Karla ; Monaghan, Kristin G. ; Muralidharan, Kasinathan ; Pratt, Victoria M. ; Prior, Thomas W. ; Richie, Kristy L. ; Levin, Barbara C. ; Rohlfs, Elizabeth M. ; Schaefer, Frederick V. ; Shrimpton, Antony E. ; Spector, Elaine B. ; Stolle, Catherine A. ; Strom, Charles M. ; Thibodeau, Stephen N. ; Cole, Eugene C. ; Goodman, Barbara K. ; Stenzel, Timothy T. / Genetically characterized positive control cell lines derived from residual clinical blood samples. In: Clinical Chemistry. 2005 ; Vol. 51, No. 11. pp. 2013-2024.
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abstract = "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 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. Methods: 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. Results: 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. Conclusions: 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.",
author = "Bernacki, {Susan H.} and Beck, {Jeanne C.} and Stankovic, {Ana K.} and Williams, {Laurina O.} and Jean Amos and Karen Snow-Bailey and Farkas, {Daniel H.} and Friez, {Michael J.} and Hantash, {Feras M.} and Karla Matteson and Monaghan, {Kristin G.} and Kasinathan Muralidharan and Pratt, {Victoria M.} and Prior, {Thomas W.} and Richie, {Kristy L.} and Levin, {Barbara C.} and Rohlfs, {Elizabeth M.} and Schaefer, {Frederick V.} and Shrimpton, {Antony E.} and Spector, {Elaine B.} and Stolle, {Catherine A.} and Strom, {Charles M.} and Thibodeau, {Stephen N.} and Cole, {Eugene C.} and Goodman, {Barbara K.} and Stenzel, {Timothy T.}",
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T1 - Genetically characterized positive control cell lines derived from residual clinical blood samples

AU - Bernacki, Susan H.

AU - Beck, Jeanne C.

AU - Stankovic, Ana K.

AU - Williams, Laurina O.

AU - Amos, Jean

AU - Snow-Bailey, Karen

AU - Farkas, Daniel H.

AU - Friez, Michael J.

AU - Hantash, Feras M.

AU - Matteson, Karla

AU - Monaghan, Kristin G.

AU - Muralidharan, Kasinathan

AU - Pratt, Victoria M.

AU - Prior, Thomas W.

AU - Richie, Kristy L.

AU - Levin, Barbara C.

AU - Rohlfs, Elizabeth M.

AU - Schaefer, Frederick V.

AU - Shrimpton, Antony E.

AU - Spector, Elaine B.

AU - Stolle, Catherine A.

AU - Strom, Charles M.

AU - Thibodeau, Stephen N.

AU - Cole, Eugene C.

AU - Goodman, Barbara K.

AU - Stenzel, Timothy T.

PY - 2005/11/1

Y1 - 2005/11/1

N2 - 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 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. Methods: 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. Results: 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. Conclusions: 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.

AB - 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 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. Methods: 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. Results: 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. Conclusions: 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.

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