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Published on: March 11, 2014
Sequence-based genotyping HPV L1 DNA and RNA transcripts in clinical specimens
Maria Satra1, Dimitra N Vamvakopoulou, Despina O Sioutopoulou
1Department of Biology and Genetics, University of Thessalia Medical School, Neo Ktirio, P.O. Box 1400, Larisa 44110, Greece. mariasatra@yahoo.com
Pathology, Research and Practice
|September 19, 2009
Summary
A new direct sequence-based genotyping method accurately identifies single and multiple human papillomavirus (HPV) L1 DNA and RNA types. This approach offers a versatile tool for HPV detection, especially for rare types and mixed infections.
Area of Science:
- Molecular Biology
- Virology
- Genetics
Background:
- Human Papillomavirus (HPV) is a common viral infection with numerous types.
- Accurate genotyping of HPV is crucial for diagnosis and management, particularly in dermatological and genital specimens.
- Existing commercial methods may have limitations in detecting rare HPV types or mixed infections.
Purpose of the Study:
- To develop and validate a direct sequence-based genotyping method for HPV L1 DNA and RNA.
- To assess the method's capability in detecting single and multiple HPV types in various specimen types.
- To establish a working cutoff value for the assay and explore its utility for rare HPV types and mixed infections.
Main Methods:
- Developed a method combining PCR amplification of a conserved HPV L1 segment with broad-spectrum primers.
- Utilized automated sequencing of PCR products followed by GenBank data sorting for genotyping.
- Validated the method on 5 skin and 30 cervical HPV DNA-positive specimens, comparing with commercial hybridization techniques.
Main Results:
- Successfully genotyped single and multiple HPV L1 DNA and RNA types in the tested specimens.
- Established the first experimentally derived working cutoff value for the sequence-based method.
- Demonstrated the method's ability to detect rare HPV types and differentiate multiple infections.
Conclusions:
- Sequence-based genotyping of amplified DNA and RNA is a viable primary method for HPV detection in dermatological samples.
- This all-purpose genotyping approach can identify rare HPV types missed by commercial techniques.
- The method facilitates the comprehensive analysis of multiple HPV infections, including prevalence ordering.
