Sentinel-base DNA genotyping using multiple sequencing primers for high-risk human papillomaviruses

• Despite the various technologies in place for genotyping human papillomaviruses (HPV), clinical use and clinical research demand a method that is fast, more reliable and cost-effective. The technology described here represents a breakthrough development in that direction. By combining the method of multiple sequencing primers with DNA sequencing, we have developed a rapid assay for genotyping HPV that relies on the identification of a single, type-specific 'sentinel' base. As described here, the prototype assay has been developed to recognize the 12 most high-risk HPV types (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59) and is capable of recognizing and simultaneously genotyping multiple HPV co-infections. By providing sequence information on multiple HPV infections, this method eliminates the need for labor- and cost-intensive PCR cloning. These proof-of-concept studies establish the assay to be accurate, reliable, rapid, flexible, and cost-effective, providing evidence of the feasibility this technique for use in clinical settings.

Subject headings

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Keyword

Cervical Intraepithelial Neoplasia/blood/diagnosis/*virology

DNA Primers/biosynthesis/*genetics

DNA Probes

HPV/genetics

DNA

Viral/blood/genetics

Female

Genotype

Humans

Papillomaviridae/classification/*genetics/isolation & purification

Papillomavirus Infections/blood/diagnosis/*virology

Polymerase Chain Reaction

Risk Factors

Sensitivity and Specificity

human papillomaviruses (HPV); DNA sequencing; multiple infections; multiple sequencing primers; sentinel-base DNA sequencing; pyrosequencing technology; POLYMERASE-CHAIN-REACTION; GENITAL HUMAN PAPILLOMAVIRUSES; CERVICAL-CANCER; HYBRID CAPTURE; GENERAL PRIMERS; NESTED PCR; SPECIMENS; SAMPLES; HPV; IDENTIFICATION

Biochemistry

Publication and Content Type

ref (subject category)

art (subject category)