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Human papillomavirus genotyping using a modified linear array detection protocol.

Matthew P Stevens1, Suzanne M Garland, Sepehr N Tabrizi

  • 1Department of Microbiology, The Royal Women's Hospital, Carlton, Vic., Australia. matthew.stevens@mcri.edu.au

Journal of Virological Methods
|March 28, 2006
PubMed
Summary
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A dry-air incubator can replace a shaking water-bath for human papillomavirus (HPV) genotyping, simplifying the assay. This modification offers near-perfect concordance, making HPV detection more accessible for laboratories.

Area of Science:

  • Molecular Biology
  • Clinical Diagnostics
  • Virology

Background:

  • A standardized PCR-based linear array assay enables concurrent typing of 37 human papillomavirus (HPV) types.
  • The assay's amplicon detection protocol traditionally requires a shaking water-bath incubator, which can be cumbersome and unavailable in many labs.

Purpose of the Study:

  • To evaluate the utility of a dry-air incubator as a substitute for the shaking water-bath in the HPV genotyping assay.
  • To determine if this modification simplifies the detection process and maintains assay accuracy.

Main Methods:

  • DNA was extracted from 150 PreservCyt specimens using the AmpliLute method.
  • PCR amplification and amplicon detection were performed using both standard (shaking water-bath) and modified (dry-air incubator) protocols.

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  • HPV-type profiles were compared between the two protocols.

    • Main Results:

    • Near-perfect concordance (kappa = 0.98) was achieved between the standard and modified protocols.
    • Identical HPV-type profiles were generated in 149 out of 150 (99.3%) specimens.
    • The dry-air incubator method demonstrated high accuracy comparable to the shaking water-bath.

    Conclusions:

    • A dry-air incubator can effectively replace a shaking water-bath in this HPV genotyping assay.
    • This modification simplifies the detection process, reduces setup time, and minimizes potential issues like water spillage.
    • The findings suggest broader adoption of this HPV detection method by laboratories after in-house validation.