HPV infection among women in a representative rural and suburban population of the USA

Abstract

DNA sequencing is a useful method for increasing the specificity of HPV genotyping as an aid to follow persistent high-risk HPV infections to reduce excessive colposcopies in populations with low cancer prevalence.

Detection of human papillomavirus (HPV) L1 gene DNA possibly bound to particulate aluminum adjuvant in the HPV vaccine Gardasil®

“…A segment of 45-60 bases of the hypervariable region of the L1 gene sequence was excised from the computer-generated base-calling electropherogram for BLAST analyses for confirmation of the HPV DNA detected and to validate its genotype. The technical detail of this nested PCR/DNA sequencing methodology has been previously reported [30][31][32][33][34][35].…”

“…Short target sequence HPV genotyping was performed by direct automated cycle DNA sequencing [30][31][32][33][34][35]. Briefly, a trace of the positive nested PCR product was transferred directly with a micro-glass rod from the positive nested PCR tube into a 20 μL volume of a cycle sequencing reaction mixture consisting of 14.5 μL water, 3.5 μL of 5 X buffer, 1 μL of BigDye Terminator 1.1 (Applied Biosystems) and 1 μL of 10 μM GP6, or GP5 sequencing primer.…”

Detection of human papillomavirus L1 gene DNA fragments in postmortem blood and spleen after Gardasil<sup>®</sup> vaccination—A case report

“…As a result, a LoTemp ® PCR with a highly processive HiFi ® DNA polymerase system programmed at thermocycling steps not to exceed 85˚C was selected for this study. The general method used to detect HPV L1 gene DNA by heminested (nested) LoTemp ® PCR amplification with the GP/MY degenerate consensus primers and validation with direct automated DNA sequenceing for genotyping has been described in detail elsewhere for clinical samples [20][21][22][23][24][25] and for detecting residual HPV DNA fragments in the Gardasil ® vaccine [9]. Each primary PCR consisted of 1 μL of reconstituted DNA solution in molecular grade water containing about 0.5 μg human DNA, 2 μL of water, 1 μL of 10 μM forward primer, 1 μL of 10 μM reverse primer, and 20 μL of ready-to-use LoTemp ® PCR master mix with HiFi ® DNA polymerase (www.hifidna.com) in a total volume of 25 μL.…”

High prevalence of HPV59 in cytologically abnormal cervical samples

“…number of lifetime male partners, age at sexual debut, bacterial infection, use of oral contraceptives, and geographic area8,10,11,12 . These independent factors are well-documented, particularly in regions where adolescents and young adults up to 29 years of age have the highest hrHPV infection rates12 .…”