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, Laurina O. Williams, Jeanne C. Beck, James E. Herndon, Karen Snow-Bailey, Thomas W. Prior, Karla Matteson, Linda M. Wasserman, Eugene C. Cole, Timothy T. Stenzel

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)227-230
Number of pages4
JournalJournal of Molecular Diagnostics
Volume5
Issue number4
DOIs
StatePublished - Jan 1 2003

Fingerprint

Transformed Cell Line
Genetic Testing
Human Herpesvirus 4
Anticoagulants
Molecular Biology
Lymphocytes
Temperature
Diamines
Hemolysis
Blood Cells
Public Health
Logistic Models
Cell Line
Acids
acid citrate dextrose
ethylene

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Molecular Medicine

Cite this

Establishment of stably EBV-transformed cell lines from residual clinical blood samples for use in performance evaluation and quality assurance in molecular genetic testing. / Bernacki, Susan H.; Stankovic, Ana K.; Williams, Laurina O.; Beck, Jeanne C.; Herndon, James E.; Snow-Bailey, Karen; Prior, Thomas W.; Matteson, Karla; Wasserman, Linda M.; Cole, Eugene C.; Stenzel, Timothy T.

In: Journal of Molecular Diagnostics, Vol. 5, No. 4, 01.01.2003, p. 227-230.

Research output: Contribution to journalArticle

Bernacki, Susan H. ; Stankovic, Ana K. ; Williams, Laurina O. ; Beck, Jeanne C. ; Herndon, James E. ; Snow-Bailey, Karen ; Prior, Thomas W. ; Matteson, Karla ; Wasserman, Linda M. ; Cole, Eugene C. ; Stenzel, Timothy T. / Establishment of stably EBV-transformed cell lines from residual clinical blood samples for use in performance evaluation and quality assurance in molecular genetic testing. In: Journal of Molecular Diagnostics. 2003 ; Vol. 5, No. 4. pp. 227-230.
@article{b34b30122769419095d742fdccdb1423,
title = "Establishment of stably EBV-transformed cell lines from residual clinical blood samples for use in performance evaluation and quality assurance in molecular genetic testing",
abstract = "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.",
author = "Bernacki, {Susan H.} and Stankovic, {Ana K.} and Williams, {Laurina O.} and Beck, {Jeanne C.} and Herndon, {James E.} and Karen Snow-Bailey and Prior, {Thomas W.} and Karla Matteson and Wasserman, {Linda M.} and Cole, {Eugene C.} and Stenzel, {Timothy T.}",
year = "2003",
month = "1",
day = "1",
doi = "10.1016/S1525-1578(10)60478-3",
language = "English (US)",
volume = "5",
pages = "227--230",
journal = "Journal of Molecular Diagnostics",
issn = "1525-1578",
publisher = "Association of Molecular Pathology",
number = "4",

}

TY - JOUR

T1 - Establishment of stably EBV-transformed cell lines from residual clinical blood samples for use in performance evaluation and quality assurance in molecular genetic testing

AU - Bernacki, Susan H.

AU - Stankovic, Ana K.

AU - Williams, Laurina O.

AU - Beck, Jeanne C.

AU - Herndon, James E.

AU - Snow-Bailey, Karen

AU - Prior, Thomas W.

AU - Matteson, Karla

AU - Wasserman, Linda M.

AU - Cole, Eugene C.

AU - Stenzel, Timothy T.

PY - 2003/1/1

Y1 - 2003/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=10744221736&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=10744221736&partnerID=8YFLogxK

U2 - 10.1016/S1525-1578(10)60478-3

DO - 10.1016/S1525-1578(10)60478-3

M3 - Article

VL - 5

SP - 227

EP - 230

JO - Journal of Molecular Diagnostics

JF - Journal of Molecular Diagnostics

SN - 1525-1578

IS - 4

ER -