Molecular Papanicolaou tests in the twenty-first century: Molecular analyses with fluid-based Papanicolaou technology

W. Michael Lin, Raheela Ashfaq, Eugenia A. Michalopulos, Anirban Maitra, Adi F. Gazdar, Carolyn Y. Muller

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

OBJECTIVE: This study was undertaken to demonstrate the feasibility of performing molecular analyses at the deoxyribonucleic acid, ribonucleic acid and protein levels of cervical cytologic examination with a methanol fluid-based Papanicolaou specimen collection system. STUDY DESIGN: Genomic deoxyribonucleic acid and total ribonucleic acid were extracted from cell pellets obtained from the residual fluid-based Papanicolaou specimen collection buffer after clinical processing. Genomic and human papillomavirus deoxyribonucleic acid polymerase chain reaction and reverse transcriptase-polymerase chain reaction were performed. Messenger ribonucleic acid transcript analysis and human papillomavirus 16 E6 mutational analysis were also performed. Methylation-specific polymerase chain reaction was used to evaluate hypermethylation status of the p16 gene and the gene for E-cadherin. Immunohistochemical staining for protein expression was performed on the processed monolayer slides. RESULTS: Cell pellets from the residual fluid-based cytologic specimen yielded good quality deoxyribonucleic acid and ribonucleic acid. Molecular analyses of genomic deoxyribonucleic acid were successful for the identification of human papillomavirus E6 and p53 polymorphism status by means of restriction enzyme digestion and direct sequencing. Methylation status of the promotor regions of the p16 tumor suppressor gene and the gene for E-cadherin were also successfully identified. Ribonucleic acid was used as the template for transcript analysis and mutational analysis of the corresponding complementary deoxyribonucleic acid of the p53 gene. Protein expression analysis was demonstrated by immunohistochemical staining for carcinoembryonic antigen. CONCLUSION: It is feasible to conduct multiple molecular analyses at the deoxyribonucleic acid, ribonucleic acid, and protein levels of the cervicovaginal cell pellets from the residual fluid-based Papanicolaou cytologic specimen. This relatively simple and widely used collection system will allow significant advances in molecular epidemiology and eventual development of a molecular Papanicolaou test.

Original languageEnglish (US)
Pages (from-to)39-45
Number of pages7
JournalAmerican Journal of Obstetrics and Gynecology
Volume183
Issue number1
DOIs
StatePublished - 2000

Fingerprint

Papanicolaou Test
Technology
RNA
DNA
Specimen Handling
Cadherins
Methylation
Proteins
Staining and Labeling
p16 Genes
Polymerase Chain Reaction
Forensic Anthropology
Human papillomavirus 16
Molecular Epidemiology
Carcinoembryonic Antigen
p53 Genes
Tumor Suppressor Genes
Reverse Transcriptase Polymerase Chain Reaction
Genetic Promoter Regions
Genes

Keywords

  • Cervical dysplasia
  • Genetic polymorphisms
  • Human papillomavirus
  • Molecular analyses

ASJC Scopus subject areas

  • Medicine(all)
  • Obstetrics and Gynecology

Cite this

Molecular Papanicolaou tests in the twenty-first century : Molecular analyses with fluid-based Papanicolaou technology. / Lin, W. Michael; Ashfaq, Raheela; Michalopulos, Eugenia A.; Maitra, Anirban; Gazdar, Adi F.; Muller, Carolyn Y.

In: American Journal of Obstetrics and Gynecology, Vol. 183, No. 1, 2000, p. 39-45.

Research output: Contribution to journalArticle

Lin, W. Michael ; Ashfaq, Raheela ; Michalopulos, Eugenia A. ; Maitra, Anirban ; Gazdar, Adi F. ; Muller, Carolyn Y. / Molecular Papanicolaou tests in the twenty-first century : Molecular analyses with fluid-based Papanicolaou technology. In: American Journal of Obstetrics and Gynecology. 2000 ; Vol. 183, No. 1. pp. 39-45.
@article{f73864ce4897457a92559b9194a9b6e6,
title = "Molecular Papanicolaou tests in the twenty-first century: Molecular analyses with fluid-based Papanicolaou technology",
abstract = "OBJECTIVE: This study was undertaken to demonstrate the feasibility of performing molecular analyses at the deoxyribonucleic acid, ribonucleic acid and protein levels of cervical cytologic examination with a methanol fluid-based Papanicolaou specimen collection system. STUDY DESIGN: Genomic deoxyribonucleic acid and total ribonucleic acid were extracted from cell pellets obtained from the residual fluid-based Papanicolaou specimen collection buffer after clinical processing. Genomic and human papillomavirus deoxyribonucleic acid polymerase chain reaction and reverse transcriptase-polymerase chain reaction were performed. Messenger ribonucleic acid transcript analysis and human papillomavirus 16 E6 mutational analysis were also performed. Methylation-specific polymerase chain reaction was used to evaluate hypermethylation status of the p16 gene and the gene for E-cadherin. Immunohistochemical staining for protein expression was performed on the processed monolayer slides. RESULTS: Cell pellets from the residual fluid-based cytologic specimen yielded good quality deoxyribonucleic acid and ribonucleic acid. Molecular analyses of genomic deoxyribonucleic acid were successful for the identification of human papillomavirus E6 and p53 polymorphism status by means of restriction enzyme digestion and direct sequencing. Methylation status of the promotor regions of the p16 tumor suppressor gene and the gene for E-cadherin were also successfully identified. Ribonucleic acid was used as the template for transcript analysis and mutational analysis of the corresponding complementary deoxyribonucleic acid of the p53 gene. Protein expression analysis was demonstrated by immunohistochemical staining for carcinoembryonic antigen. CONCLUSION: It is feasible to conduct multiple molecular analyses at the deoxyribonucleic acid, ribonucleic acid, and protein levels of the cervicovaginal cell pellets from the residual fluid-based Papanicolaou cytologic specimen. This relatively simple and widely used collection system will allow significant advances in molecular epidemiology and eventual development of a molecular Papanicolaou test.",
keywords = "Cervical dysplasia, Genetic polymorphisms, Human papillomavirus, Molecular analyses",
author = "Lin, {W. Michael} and Raheela Ashfaq and Michalopulos, {Eugenia A.} and Anirban Maitra and Gazdar, {Adi F.} and Muller, {Carolyn Y.}",
year = "2000",
doi = "10.1067/mob.2000.105734",
language = "English (US)",
volume = "183",
pages = "39--45",
journal = "American Journal of Obstetrics and Gynecology",
issn = "0002-9378",
publisher = "Mosby Inc.",
number = "1",

}

TY - JOUR

T1 - Molecular Papanicolaou tests in the twenty-first century

T2 - Molecular analyses with fluid-based Papanicolaou technology

AU - Lin, W. Michael

AU - Ashfaq, Raheela

AU - Michalopulos, Eugenia A.

AU - Maitra, Anirban

AU - Gazdar, Adi F.

AU - Muller, Carolyn Y.

PY - 2000

Y1 - 2000

N2 - OBJECTIVE: This study was undertaken to demonstrate the feasibility of performing molecular analyses at the deoxyribonucleic acid, ribonucleic acid and protein levels of cervical cytologic examination with a methanol fluid-based Papanicolaou specimen collection system. STUDY DESIGN: Genomic deoxyribonucleic acid and total ribonucleic acid were extracted from cell pellets obtained from the residual fluid-based Papanicolaou specimen collection buffer after clinical processing. Genomic and human papillomavirus deoxyribonucleic acid polymerase chain reaction and reverse transcriptase-polymerase chain reaction were performed. Messenger ribonucleic acid transcript analysis and human papillomavirus 16 E6 mutational analysis were also performed. Methylation-specific polymerase chain reaction was used to evaluate hypermethylation status of the p16 gene and the gene for E-cadherin. Immunohistochemical staining for protein expression was performed on the processed monolayer slides. RESULTS: Cell pellets from the residual fluid-based cytologic specimen yielded good quality deoxyribonucleic acid and ribonucleic acid. Molecular analyses of genomic deoxyribonucleic acid were successful for the identification of human papillomavirus E6 and p53 polymorphism status by means of restriction enzyme digestion and direct sequencing. Methylation status of the promotor regions of the p16 tumor suppressor gene and the gene for E-cadherin were also successfully identified. Ribonucleic acid was used as the template for transcript analysis and mutational analysis of the corresponding complementary deoxyribonucleic acid of the p53 gene. Protein expression analysis was demonstrated by immunohistochemical staining for carcinoembryonic antigen. CONCLUSION: It is feasible to conduct multiple molecular analyses at the deoxyribonucleic acid, ribonucleic acid, and protein levels of the cervicovaginal cell pellets from the residual fluid-based Papanicolaou cytologic specimen. This relatively simple and widely used collection system will allow significant advances in molecular epidemiology and eventual development of a molecular Papanicolaou test.

AB - OBJECTIVE: This study was undertaken to demonstrate the feasibility of performing molecular analyses at the deoxyribonucleic acid, ribonucleic acid and protein levels of cervical cytologic examination with a methanol fluid-based Papanicolaou specimen collection system. STUDY DESIGN: Genomic deoxyribonucleic acid and total ribonucleic acid were extracted from cell pellets obtained from the residual fluid-based Papanicolaou specimen collection buffer after clinical processing. Genomic and human papillomavirus deoxyribonucleic acid polymerase chain reaction and reverse transcriptase-polymerase chain reaction were performed. Messenger ribonucleic acid transcript analysis and human papillomavirus 16 E6 mutational analysis were also performed. Methylation-specific polymerase chain reaction was used to evaluate hypermethylation status of the p16 gene and the gene for E-cadherin. Immunohistochemical staining for protein expression was performed on the processed monolayer slides. RESULTS: Cell pellets from the residual fluid-based cytologic specimen yielded good quality deoxyribonucleic acid and ribonucleic acid. Molecular analyses of genomic deoxyribonucleic acid were successful for the identification of human papillomavirus E6 and p53 polymorphism status by means of restriction enzyme digestion and direct sequencing. Methylation status of the promotor regions of the p16 tumor suppressor gene and the gene for E-cadherin were also successfully identified. Ribonucleic acid was used as the template for transcript analysis and mutational analysis of the corresponding complementary deoxyribonucleic acid of the p53 gene. Protein expression analysis was demonstrated by immunohistochemical staining for carcinoembryonic antigen. CONCLUSION: It is feasible to conduct multiple molecular analyses at the deoxyribonucleic acid, ribonucleic acid, and protein levels of the cervicovaginal cell pellets from the residual fluid-based Papanicolaou cytologic specimen. This relatively simple and widely used collection system will allow significant advances in molecular epidemiology and eventual development of a molecular Papanicolaou test.

KW - Cervical dysplasia

KW - Genetic polymorphisms

KW - Human papillomavirus

KW - Molecular analyses

UR - http://www.scopus.com/inward/record.url?scp=0033855555&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033855555&partnerID=8YFLogxK

U2 - 10.1067/mob.2000.105734

DO - 10.1067/mob.2000.105734

M3 - Article

C2 - 10920306

AN - SCOPUS:0033855555

VL - 183

SP - 39

EP - 45

JO - American Journal of Obstetrics and Gynecology

JF - American Journal of Obstetrics and Gynecology

SN - 0002-9378

IS - 1

ER -