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.
Gharizadeh
Baback
B
Stanford Genome Technology Center, Stanford University, 855 California Avenue, Palo Alto, CA 94304, USA.
Zheng
Biying
B
Akhras
Michael
M
Ghaderi
Mehran
M
Jejelowo
Olufisayo
O
Strander
Björn
B
Nyrén
Pål
P
Wallin
Keng-Ling
KL
Pourmand
Nader
N
D017931:DNA Primers / Q000096:biosynthesis / Q000235:genetics*; D015344:DNA Probes, HPV / Q000235:genetics; D004279:DNA, Viral / Q000097:blood / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000097:blood / Q000175:diagnosis / Q000821:virology*; D016133:Polymerase Chain Reaction; D012307:Risk Factors; D012680:Sensitivity and Specificity; D002578:Uterine Cervical Dysplasia / Q000097:blood / Q000175:diagnosis / Q000821:virology*
D017931:DNA Primers; D015344:DNA Probes, HPV; D004279:DNA, Viral