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Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
Bead-based multiplex genotyping of 58 cutaneous human papillomavirus types
K M Michael1, O Forslund, O Bacevskij
1Department of Genome Modifications and Carcinogenesis, Infection and Cancer Program, German Cancer Research Center, 69120 Heidelberg, Germany.
Journal of Clinical Microbiology
|August 12, 2011
Summary
A new multiplex assay (McPG) efficiently genotypes numerous cutaneous human papillomaviruses (HPVs) from skin samples. This high-throughput method improves detection of multiple HPV infections, aiding large-scale skin cancer studies.
Area of Science:
- Virology
- Dermatology
- Molecular Biology
Background:
- Cutaneous human papillomaviruses (HPVs) are diverse, with over 50 types, and their role in skin cancer is unclear.
- Existing PCR methods for genotyping cutaneous HPVs are complex and not suitable for high-throughput analysis.
- Accurate and efficient genotyping is crucial for understanding HPV natural history and disease association.
Purpose of the Study:
- To develop and validate a novel multiplex assay for high-throughput genotyping of cutaneous HPVs.
- To assess the performance of the multiplex cutaneous papillomavirus genotyping (McPG) assay.
Main Methods:
- Developed a multiplex cutaneous papillomavirus genotyping (McPG) assay using modified FAP PCR and Luminex bead-based technology.
- Utilized type-specific oligonucleotide probes for detecting 38 defined and 20 putative cutaneous HPVs across beta, gamma, mu, and nu genera.
- Incorporated β-globin primers for simultaneous DNA quality control.
Main Results:
- The McPG assay demonstrated high reproducibility (κ=0.84) and analytical sensitivity (≤100 genome copies).
- Concordance with nested FAP PCR followed by sequencing was 70.2% for 322 skin biopsy samples.
- The assay improved detection of multiple HPV infections (average 2.5 types/sample vs. 1.7 types/sample).
Conclusions:
- The McPG assay is a powerful, high-throughput tool for genotyping multiple cutaneous HPVs.
- This assay is suitable for large-scale epidemiological studies investigating HPV and skin cancer.
- McPG offers improved efficiency and accuracy for cutaneous HPV detection.
