Change search
ReferencesLink to record
Permanent link

Direct link
Type-specific multiple sequencing primers - A novel strategy for reliable and rapid genotyping of human papilloma viruses by pyrosequencing technology
Show others and affiliations
2005 (English)In: Journal of Molecular Diagnostics, ISSN 1525-1578, E-ISSN 1943-7811, Vol. 7, no 2, 198-205 p.Article in journal (Refereed) Published
Abstract [en]

DNA sequencing is the gold standard method for accurate microbial and viral typing. However, DNA sequencing techniques have been facing limitations in typing of human papillomaviruses when the specimen harbors multiple genotypes and yields nonspecific amplification products, resulting in nonspecific and noninterpretable sequence data. To address these limitations we have developed a type-specific multiple sequencing primer DNA-sequencing method. This new strategy is suitable for sequencing and typing of samples harboring different genotypes (co-infections with multiple genotypes) and yielding nonspecific amplifications, thus eliminating the need for nested polymerase chain reaction (PCR), stringent PCR conditions, and cloning. The new approach has also proved useful for amplicons containing low PCR yield or subdominant types, avoiding reperforming of amplifications. We have applied the multiple sequencing primer method for genotyping of clinically relevant human papillomaviruses in a clinical test panel by using a combined pool of seven type-specific sequencing primers for HPV-6, -11, -16, -18, -31, -33, and -45. Furthermore, we introduced a sequence pattern recognition approach when there was a plurality of genotypes in the sample to facilitate typing of more than one target DNA in the sample. The multiple sequencing primer method has proved to be a multifaceted approach for typing of human papillomaviruses by DNA sequencing technologies.

Place, publisher, year, edition, pages
2005. Vol. 7, no 2, 198-205 p.
Keyword [en]
invasive cervical-cancer, worldwide, dna, infection, lesions
URN: urn:nbn:se:kth:diva-14706ISI: 000228736900007ScopusID: 2-s2.0-18744396366OAI: diva2:332747
QC 20100525Available from: 2010-08-05 Created: 2010-08-05Bibliographically approved

Open Access in DiVA

No full text


Search in DiVA

By author/editor
Ahmadian, AfshinNyrén, Pål
By organisation
Gene TechnologyBiochemistry
In the same journal
Journal of Molecular Diagnostics

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Total: 17 hits
ReferencesLink to record
Permanent link

Direct link