Current Microbiology

, Volume 52, Issue 3, pp 204–209

Oligonucleotide Microarray with RD-PCR Labeling Technique for Detection and Typing of Human Papillomavirus


  • Wei Min
    • Institute of Molecular BiologySouthern Medical University
    • Institute of Molecular BiologySouthern Medical University
  • Zhang Bao
    • Institute of Molecular BiologySouthern Medical University
  • Li Ling
    • Institute of Molecular BiologySouthern Medical University
  • Sun Zhao-hui
    • Institute of Molecular BiologySouthern Medical University
  • Zheng Wen-ling
    • Southern China Genomics Research Center

DOI: 10.1007/s00284-005-0212-x

Cite this article as:
Min, W., Wen-li, M., Bao, Z. et al. Curr Microbiol (2006) 52: 204. doi:10.1007/s00284-005-0212-x


Currently, screening for high-risk human papillomavirus (HPV) infection remains an important health concern throughout the world, because of the close association between certain types of HPV and cervical cancer. In this study, we explore the possibility of using ∼70mer oligonucleotide microarray for detection and genotyping of HPV. The ∼70mer type-specific oligonucleotide probes of four different types HPV were designed by using biological software Arraydesigner 2.0, which analyzed the whole genome sequences of HPV and selected optimal probes. These probes were synthesized and printed onto the surface of glass slides in order to prepare a low-density microarray. HPV samples were labeled with fluorescence dyes Cy3 using a method of restriction display polymerase chain reaction (RD-PCR). HPV plasmid DNA was restricted with Sau3A I to produce multiple fragments that were ligated to adaptors subsequently and used as PCR template. PCR labeling was performed with the fluorescently labeled universal primer (Cy3-UP) whose sequence is designed according to the adaptor of the RD-PCR approaches. The labeled samples were hybridized with the oligonucleotide microarray. The scanning results showed that HPV DNA hybridized specifically with multiple spots correspondingly to show positive signals, whereas no signals were detected of all the negative and blank controls. These results demonstrated that ∼70mer oligonucleotide microarray can be applied to HPV detection and genotyping. The application of RD-PCR in the sample labeling can increase significantly the sensitivity of the assay and will be especially useful for the discriminate diagnosis of multiple pathogens.

Copyright information

© Springer Science+Business Media, Inc. 2006