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DNA microarray format for detection and subtyping of human papillomavirus
Corné H W Klaassen1, Clemens F M Prinsen, Hanneke A de Valk
1Department of Pathology, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands. c.klassen@cwz.nl
Journal of Clinical Microbiology
|May 8, 2004
Summary
A novel human papillomavirus (HPV) assay utilizes DNA microarrays for rapid detection and identification of 53 mucosal HPV types. This accessible method aids diagnostic laboratories with PCR capabilities in identifying single and multiple HPV infections.
Area of Science:
- Molecular Biology
- Virology
- Diagnostics
Background:
- Human papillomavirus (HPV) infections are a major cause of cervical cancer.
- Accurate and accessible diagnostic methods for HPV detection are crucial for effective screening and management.
Purpose of the Study:
- To describe a new human papillomavirus (HPV) assay utilizing high-density DNA microarrays.
- To enable rapid and accurate identification of a broad range of mucosal HPV types.
Main Methods:
- Amplification and digoxigenin labeling of an HPV DNA fragment (E1 gene) via PCR.
- Hybridization of amplicons to type-specific oligonucleotides on high-density DNA microarrays.
- Detection using immunohistochemical staining with a dual colorimetric and fluorescent substrate.
Main Results:
- The assay successfully detected and identified 53 mucosal HPV types.
- 45 HPV types were identified by single probes, and 8 by probe combinations.
- The method allows for rapid spot identification via light microscopy and semiquantification via laser scanning.
Conclusions:
- This HPV assay provides a simple, accessible, and effective method for identifying multiple HPV types.
- The assay's compatibility with standard PCR facilities makes it suitable for widespread diagnostic use.
- It facilitates the identification of both single and mixed HPV infections.