Stability of archived liquid-based cervical cytologic specimens

Philip E. Castle, Diane Solomon, Allan Hildesheim, Rolando Herrero, M. Concepcion Bratti, Mark E. Sherman, Ana Cecilia Rodriguez, Mario Alfaro, Martha L. Hutchinson, S. Terence Dunn, Jane Kuypers, Mark Schiffman

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Abstract

BACKGROUND. Exfoliated cervical cell specimens collected in PreservCyt®, a methanol-based medium used in ThinPrep® liquid-based cytology, have been archived in epidemiologic studies. However, long-term DNA stability and cytologic stability of these biospecimens have not been evaluated. METHODS. Cervical specimens were collected into PreservCyt from participants in a natural history study of human papillomavirus (HPV) infection and cervical carcinoma in Guanacaste, Costa Rica (1993-2000), and stored at ambient temperatures. Thirty specimens classified as low-grade squamous intraepithelial lesions by liquid-based cytology were randomly chosen from each collection year (except for 1994) and selectively assessed for molecular and cytologic stability. Specimens were tested in 2001 for 1) HPV DNA by the Hybrid Capture 2 test, 2) β-globin DNA by polymerase chain reaction amplification of multiple length fragments (268, 610, and 1327 bp), and 3) nuclear preservation by visual inspection of newly made liquid-based cytology slides. All testing was done masked to year of collection. Associations of stability and storage time were evaluated using standard contingency tables and chi-square tests for trend. RESULTS. Human papillomavirus DNA, as detected by the Hybrid Capture 2 test, was unaffected by storage time. Stability of β-globin DNA (PTrend < 0.0001) and nuclear preservation (PTrend < 0.0001) declined with increasing storage time. Approximately 15% of specimens could not be amplified for any β-globin DNA fragment after 5 years of storage (collected in 1996). In addition, cytology slides made from 41% specimens were rated as marginal (32%) or unsatisfactory (9%) after 8 years of storage (collected in 1993). CONCLUSIONS. Cervical specimens archived in PreservCyt underwent partial DNA and cytologic degradation after several years of storage. Methodologic studies to optimize long-term storage of cervical cells for epidemiologic studies of cervical carcinoma are needed.

Original languageEnglish (US)
Pages (from-to)89-96
Number of pages8
JournalCancer
Volume99
Issue number2
DOIs
StatePublished - Apr 25 2003

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Globins
Cell Biology
DNA
Epidemiologic Studies
Carcinoma
Costa Rica
Papillomavirus Infections
DNA-Directed DNA Polymerase
Chi-Square Distribution
Natural History
Methanol
Polymerase Chain Reaction
Temperature

Keywords

  • Archival
  • DNA
  • HPV
  • Liquid-based cytology
  • Stability

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Castle, P. E., Solomon, D., Hildesheim, A., Herrero, R., Bratti, M. C., Sherman, M. E., ... Schiffman, M. (2003). Stability of archived liquid-based cervical cytologic specimens. Cancer, 99(2), 89-96. https://doi.org/10.1002/cncr.11058

Stability of archived liquid-based cervical cytologic specimens. / Castle, Philip E.; Solomon, Diane; Hildesheim, Allan; Herrero, Rolando; Bratti, M. Concepcion; Sherman, Mark E.; Rodriguez, Ana Cecilia; Alfaro, Mario; Hutchinson, Martha L.; Dunn, S. Terence; Kuypers, Jane; Schiffman, Mark.

In: Cancer, Vol. 99, No. 2, 25.04.2003, p. 89-96.

Research output: Contribution to journalArticle

Castle, PE, Solomon, D, Hildesheim, A, Herrero, R, Bratti, MC, Sherman, ME, Rodriguez, AC, Alfaro, M, Hutchinson, ML, Dunn, ST, Kuypers, J & Schiffman, M 2003, 'Stability of archived liquid-based cervical cytologic specimens', Cancer, vol. 99, no. 2, pp. 89-96. https://doi.org/10.1002/cncr.11058
Castle PE, Solomon D, Hildesheim A, Herrero R, Bratti MC, Sherman ME et al. Stability of archived liquid-based cervical cytologic specimens. Cancer. 2003 Apr 25;99(2):89-96. https://doi.org/10.1002/cncr.11058
Castle, Philip E. ; Solomon, Diane ; Hildesheim, Allan ; Herrero, Rolando ; Bratti, M. Concepcion ; Sherman, Mark E. ; Rodriguez, Ana Cecilia ; Alfaro, Mario ; Hutchinson, Martha L. ; Dunn, S. Terence ; Kuypers, Jane ; Schiffman, Mark. / Stability of archived liquid-based cervical cytologic specimens. In: Cancer. 2003 ; Vol. 99, No. 2. pp. 89-96.
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abstract = "BACKGROUND. Exfoliated cervical cell specimens collected in PreservCyt{\circledR}, a methanol-based medium used in ThinPrep{\circledR} liquid-based cytology, have been archived in epidemiologic studies. However, long-term DNA stability and cytologic stability of these biospecimens have not been evaluated. METHODS. Cervical specimens were collected into PreservCyt from participants in a natural history study of human papillomavirus (HPV) infection and cervical carcinoma in Guanacaste, Costa Rica (1993-2000), and stored at ambient temperatures. Thirty specimens classified as low-grade squamous intraepithelial lesions by liquid-based cytology were randomly chosen from each collection year (except for 1994) and selectively assessed for molecular and cytologic stability. Specimens were tested in 2001 for 1) HPV DNA by the Hybrid Capture 2 test, 2) β-globin DNA by polymerase chain reaction amplification of multiple length fragments (268, 610, and 1327 bp), and 3) nuclear preservation by visual inspection of newly made liquid-based cytology slides. All testing was done masked to year of collection. Associations of stability and storage time were evaluated using standard contingency tables and chi-square tests for trend. RESULTS. Human papillomavirus DNA, as detected by the Hybrid Capture 2 test, was unaffected by storage time. Stability of β-globin DNA (PTrend < 0.0001) and nuclear preservation (PTrend < 0.0001) declined with increasing storage time. Approximately 15{\%} of specimens could not be amplified for any β-globin DNA fragment after 5 years of storage (collected in 1996). In addition, cytology slides made from 41{\%} specimens were rated as marginal (32{\%}) or unsatisfactory (9{\%}) after 8 years of storage (collected in 1993). CONCLUSIONS. Cervical specimens archived in PreservCyt underwent partial DNA and cytologic degradation after several years of storage. Methodologic studies to optimize long-term storage of cervical cells for epidemiologic studies of cervical carcinoma are needed.",
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AU - Sherman, Mark E.

AU - Rodriguez, Ana Cecilia

AU - Alfaro, Mario

AU - Hutchinson, Martha L.

AU - Dunn, S. Terence

AU - Kuypers, Jane

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N2 - BACKGROUND. Exfoliated cervical cell specimens collected in PreservCyt®, a methanol-based medium used in ThinPrep® liquid-based cytology, have been archived in epidemiologic studies. However, long-term DNA stability and cytologic stability of these biospecimens have not been evaluated. METHODS. Cervical specimens were collected into PreservCyt from participants in a natural history study of human papillomavirus (HPV) infection and cervical carcinoma in Guanacaste, Costa Rica (1993-2000), and stored at ambient temperatures. Thirty specimens classified as low-grade squamous intraepithelial lesions by liquid-based cytology were randomly chosen from each collection year (except for 1994) and selectively assessed for molecular and cytologic stability. Specimens were tested in 2001 for 1) HPV DNA by the Hybrid Capture 2 test, 2) β-globin DNA by polymerase chain reaction amplification of multiple length fragments (268, 610, and 1327 bp), and 3) nuclear preservation by visual inspection of newly made liquid-based cytology slides. All testing was done masked to year of collection. Associations of stability and storage time were evaluated using standard contingency tables and chi-square tests for trend. RESULTS. Human papillomavirus DNA, as detected by the Hybrid Capture 2 test, was unaffected by storage time. Stability of β-globin DNA (PTrend < 0.0001) and nuclear preservation (PTrend < 0.0001) declined with increasing storage time. Approximately 15% of specimens could not be amplified for any β-globin DNA fragment after 5 years of storage (collected in 1996). In addition, cytology slides made from 41% specimens were rated as marginal (32%) or unsatisfactory (9%) after 8 years of storage (collected in 1993). CONCLUSIONS. Cervical specimens archived in PreservCyt underwent partial DNA and cytologic degradation after several years of storage. Methodologic studies to optimize long-term storage of cervical cells for epidemiologic studies of cervical carcinoma are needed.

AB - BACKGROUND. Exfoliated cervical cell specimens collected in PreservCyt®, a methanol-based medium used in ThinPrep® liquid-based cytology, have been archived in epidemiologic studies. However, long-term DNA stability and cytologic stability of these biospecimens have not been evaluated. METHODS. Cervical specimens were collected into PreservCyt from participants in a natural history study of human papillomavirus (HPV) infection and cervical carcinoma in Guanacaste, Costa Rica (1993-2000), and stored at ambient temperatures. Thirty specimens classified as low-grade squamous intraepithelial lesions by liquid-based cytology were randomly chosen from each collection year (except for 1994) and selectively assessed for molecular and cytologic stability. Specimens were tested in 2001 for 1) HPV DNA by the Hybrid Capture 2 test, 2) β-globin DNA by polymerase chain reaction amplification of multiple length fragments (268, 610, and 1327 bp), and 3) nuclear preservation by visual inspection of newly made liquid-based cytology slides. All testing was done masked to year of collection. Associations of stability and storage time were evaluated using standard contingency tables and chi-square tests for trend. RESULTS. Human papillomavirus DNA, as detected by the Hybrid Capture 2 test, was unaffected by storage time. Stability of β-globin DNA (PTrend < 0.0001) and nuclear preservation (PTrend < 0.0001) declined with increasing storage time. Approximately 15% of specimens could not be amplified for any β-globin DNA fragment after 5 years of storage (collected in 1996). In addition, cytology slides made from 41% specimens were rated as marginal (32%) or unsatisfactory (9%) after 8 years of storage (collected in 1993). CONCLUSIONS. Cervical specimens archived in PreservCyt underwent partial DNA and cytologic degradation after several years of storage. Methodologic studies to optimize long-term storage of cervical cells for epidemiologic studies of cervical carcinoma are needed.

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