Semiautomated typing of human papillomaviruses by restriction fragment length polymorphism analysis of fluorescence-labeled PCR fragments

Tian Shiang Wang, Huar Fen Chou, Wei Ming Liu, Kong Bung Choo

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A simplified version of a PCR-based reductional restriction fragment length polymorphism (rRFLP) approach for typing of human papillomaviruses (HPVs) is described previously [Wang et al., 1997]. It is achieved by the use of a biotin-labeled primer in PCR which, on restriction digestion and staining, is associated with only a single restriction fragment. In this report, we describe a further development of the rRFLP approach with the use of a fluorescence-labeled primer in PCR and fragment detection by laser scanning in an automatic sequencer. HPV typing is achieved by computer- assisted matching of the fluorescence-labeled rRFLP patterns with a database of rRFLP patterns of all known anogenital HPV types. On analysis of the typing of 133 HPV-positive cases using this procedure, 20 different HPV types were detected in exfoliated cervical cells in PAP smear samples derived from Taiwanese women. The results indicate the existence of a heterogeneous population of HPV types in Taiwan. Although most cases were associated with the more common HPV types, a significant fraction (about 20%) of the HPV types detected was related to the less common genotypes, which are often not included in commercial kits available for HPV typing. The results indicate the importance of covering as many HPV types as possible in clinical HPV genotyping protocols.

Original languageEnglish
Pages (from-to)536-540
Number of pages5
JournalJournal of Medical Virology
Volume59
Issue number4
DOIs
Publication statusPublished - 1999
Externally publishedYes

Fingerprint

Restriction Fragment Length Polymorphisms
Fluorescence
Polymerase Chain Reaction
Biotin
Taiwan
Digestion
Lasers
Genotype
Databases
Staining and Labeling

Keywords

  • Fluorescence labeling
  • Human papillomavirus
  • RFLP
  • Typing

ASJC Scopus subject areas

  • Virology

Cite this

Semiautomated typing of human papillomaviruses by restriction fragment length polymorphism analysis of fluorescence-labeled PCR fragments. / Wang, Tian Shiang; Chou, Huar Fen; Liu, Wei Ming; Choo, Kong Bung.

In: Journal of Medical Virology, Vol. 59, No. 4, 1999, p. 536-540.

Research output: Contribution to journalArticle

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