Simultaneous amplification and identification of 25 human papillomavirus types with Templex technology.
The majority of existing human papillomavirus (HPV) genotyping assays are based on multiplex PCR using consensus or degenerate primers. We developed a Templex HPV assay that simultaneously detects and identifies 25 common HPV genotypes in a single-tube reaction using type-specific primers for the HPV-specific E6 and E7 genes. The analytical sensitivities of the Templex assay for HPV type 16 (HPV-16), -18, and -56 were 20, 100, and 20 copies per reaction mixture, respectively. The Templex assay provides semiquantitative information on each type when multiple HPV types coexist in one reaction. We tested 109 clinical cervical specimens previously evaluated with the Digene HC2 high-risk HPV DNA test and found 95.4% concordance between the assay results. The Templex assay provided type-specific results and found multiple types in 29.2% (14 of 48) of high-risk HPV-positive samples. The entire Templex procedure, including DNA extraction, can be completed within 5 hours, providing a rapid and reliable diagnostic tool for HPV detection and typing that is amenable to automation.
Han
Jian
J
Genaco Biomedical Products, Inc., Huntsville, Alabama 35805, USA.
Swan
David C
DC
Smith
Sharon J
SJ
Lum
Shanjuan H
SH
Sefers
Susan E
SE
Unger
Elizabeth R
ER
Tang
Yi-Wei
YW
D002584:Cervix Uteri / Q000821:virology; D005260:Female; D005838:Genotype; D052162:Human papillomavirus 16 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D052161:Human papillomavirus 18 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010957:Plasmids; D016133:Polymerase Chain Reaction