TY - JOUR
T1 - Human papillomavirus genotyping after denaturation of specimens for Hybrid Capture 2 testing
T2 - Feasibility study for the HPV persistence and progression cohort
AU - LaMere, Brandon J.
AU - Kornegay, Janet
AU - Fetterman, Barbara
AU - Sadorra, Mark
AU - Shieh, Jen
AU - Castle, Philip E.
N1 - Funding Information:
This research was supported by the Intramural Research Program of the NIH, NCI and by Kaiser Permanente Northern California (KPNC). We thank Dr. Gene Pawlick, former director of the KPNC Clinical Lab, for his unwavering support of this project.
PY - 2007/12
Y1 - 2007/12
N2 - Human papillomavirus (HPV) genotyping could be clinically useful, depending on the results of large, prospective studies like the HPV persistence and progression (PaP) cohort. The cohort is based on genotyping and follow-up of Hybrid Capture-positive women at Kaiser Permanente, Northern California. HPV DNA testing by Hybrid Capture 2 requires denaturation with alkali, possibly damaging the DNA for optimal PCR-based genotyping. A feasibility study was conducted on paired aliquots of anonymized specimens from 100 women with low-grade intraepithelial lesion cytology. Test aliquots were left in denaturant for 10 or 18 h at 4 °C and then neutralized; comparison aliquots were not denatured but diluted to match the timing, temperature, concentration and salt conditions of the treated specimens. The masked aliquots were tested using a commercialized PCR-based assay that detects of 37 HPV genotypes. There was no overall effect of treatment on test positivity or number of types. HPV16 was marginally more likely to be detected in untreated versus treated aliquots (P = 0.09) but HPV45 was marginally more likely to be detected in treated than untreated aliquots (P = 0.07), suggesting that these differences represented chance (intra-test variability). It can be concluded that residual Hybrid Capture-positive specimens can be genotyped by PCR after Hybrid Capture 2 processing.
AB - Human papillomavirus (HPV) genotyping could be clinically useful, depending on the results of large, prospective studies like the HPV persistence and progression (PaP) cohort. The cohort is based on genotyping and follow-up of Hybrid Capture-positive women at Kaiser Permanente, Northern California. HPV DNA testing by Hybrid Capture 2 requires denaturation with alkali, possibly damaging the DNA for optimal PCR-based genotyping. A feasibility study was conducted on paired aliquots of anonymized specimens from 100 women with low-grade intraepithelial lesion cytology. Test aliquots were left in denaturant for 10 or 18 h at 4 °C and then neutralized; comparison aliquots were not denatured but diluted to match the timing, temperature, concentration and salt conditions of the treated specimens. The masked aliquots were tested using a commercialized PCR-based assay that detects of 37 HPV genotypes. There was no overall effect of treatment on test positivity or number of types. HPV16 was marginally more likely to be detected in untreated versus treated aliquots (P = 0.09) but HPV45 was marginally more likely to be detected in treated than untreated aliquots (P = 0.07), suggesting that these differences represented chance (intra-test variability). It can be concluded that residual Hybrid Capture-positive specimens can be genotyped by PCR after Hybrid Capture 2 processing.
KW - Cervical cancer
KW - Cervical intraepithelial neoplasia grade 3 (CIN3)
KW - Cervical precancer
KW - HPV genotypes
KW - Human papillomavirus (HPV)
KW - PCR
KW - Screening
UR - http://www.scopus.com/inward/record.url?scp=35649017206&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=35649017206&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2007.06.001
DO - 10.1016/j.jviromet.2007.06.001
M3 - Article
C2 - 17673302
AN - SCOPUS:35649017206
SN - 0166-0934
VL - 146
SP - 80
EP - 85
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1-2
ER -