title: A novel real-time genotyping assay for detection of the E6-350G HPV 16 variant.
abstract: It has been suggested that some E6 human papillomavirus (HPV) type 16 variants could be involved in viral persistence and progression of HPV infection. A novel one-step allelic discrimination real-time PCR was evaluated for E6-350G variant detection in 102 endocervical HPV 16 positive samples. This assay was also used to assess the distribution of this variant in Spanish women with cervical cancer related to HPV 16. The detection limit for the allelic discrimination assay was 50 copies per reaction, even where the E6-350G variant represents only 20% of the variants in the sample. Complete concordance was observed between DNA sequencing and the novel AD RT-PCR assay. Fourteen E6-350T reference strains and 18 E6-350G variants were detected out of 32 endocervical samples from women with cervical cancer. The average age of women who were infected by the E6-350G HPV 16 variant was 10 years lower in these samples than in women who were infected by the reference strain. This novel allelic discrimination assay is a fast, sensitive and specific method for detection of the E6-350G HPV 16 variant.
authors.affiliation: Department of Microbiology, Complexo Hospitalario Universitario de Vigo, Vigo, Spain. sonia.perez.castro@sergas.es
authors.forename: S
authors.initials: S
authors.lastname: Perez
authors.forename: A
authors.initials: A
authors.lastname: Cid
authors.forename: A
authors.initials: A
authors.lastname: Araujo
authors.forename: M J
authors.initials: MJ
authors.lastname: Lamas
authors.forename: M T
authors.initials: MT
authors.lastname: Saran
authors.forename: M J
authors.initials: MJ
authors.lastname: Alvarez
authors.forename: I
authors.initials: I
authors.lastname: Lopez-Miragaya
authors.forename: S
authors.initials: S
authors.lastname: Gonzalez
authors.forename: J
authors.initials: J
authors.lastname: Torres
authors.forename: S
authors.initials: S
authors.lastname: Melon
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000483:Alleles; D005260:Female; D005838:Genotype; D052162:Human papillomavirus 16 / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008875:Middle Aged; D058889:Molecular Typing / Q000379:methods*; D020125:Mutation, Missense; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D030361:Papillomavirus Infections / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012097:Repressor Proteins / Q000235:genetics*; D012680:Sensitivity and Specificity; D013030:Spain; D055815:Young Adult
chemical_list: C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins