TY - JOUR
T1 - Automated high throughput DNA isolation for detection of human papillomavirus in oral rinse samples
AU - Broutian, Tatevik R.
AU - He, Xin
AU - Gillison, Maura L.
N1 - Funding Information:
This study was funded by a grant (DE016631) from the National Institute of Dental and Craniofacial Research (NIDCR).
PY - 2011/4
Y1 - 2011/4
N2 - Background: Oral HPV infection elevates risk of oropharyngeal cancer, but its natural history is unknown. Natural history studies necessitate validation of an automated, high-throughput method for HPV genomic DNA detection in oral rinse samples (ORS). Objectives: To compare agreement of oral HPV detection in ORS processed by a magnetic-bead based automated platform to a previous gold-standard, manual protein-precipitation method. Agreement was compared to that of repeat sampling and repeat HPV testing. Study design: HIV-infected individuals (n = 100) provided two ORS collected 15. min apart. DNA was isolated from equal aliquots by either a protein-precipitation based (Puregene, Qiagen) or magnetic bead-based (QIAsymphony™ SP, Qiagen) method. HPV DNA was detected and type-specified by consensus primer PCR and reverse line blot hybridization. The kappa statistic was used to assess overall agreement (OA) and agreement on a positive test (Ps+). Results: The DNA purification methods had very high agreement for categorizing an individual as HPV infected (OA. = 0.95; Ps+. = 0.94) as well as for detection of HPV type-specific infection (OA. = 0.99; Ps+. = 0.88) in ORS. Agreement for detection of HPV type-specific infection was greater than that observed with repeat oral rinse sampling (OA. = 0.99, Ps+. = 0.76) but comparable to inter-assay agreement (OA. = 1.00, Ps+. = 0.90). Conclusions: HPV detection in ORS processed with a magnetic-bead based automated platform will facilitate large natural history studies of oral HPV infection necessary to evaluate the potential use of oral HPV detection in oral cancer screening.
AB - Background: Oral HPV infection elevates risk of oropharyngeal cancer, but its natural history is unknown. Natural history studies necessitate validation of an automated, high-throughput method for HPV genomic DNA detection in oral rinse samples (ORS). Objectives: To compare agreement of oral HPV detection in ORS processed by a magnetic-bead based automated platform to a previous gold-standard, manual protein-precipitation method. Agreement was compared to that of repeat sampling and repeat HPV testing. Study design: HIV-infected individuals (n = 100) provided two ORS collected 15. min apart. DNA was isolated from equal aliquots by either a protein-precipitation based (Puregene, Qiagen) or magnetic bead-based (QIAsymphony™ SP, Qiagen) method. HPV DNA was detected and type-specified by consensus primer PCR and reverse line blot hybridization. The kappa statistic was used to assess overall agreement (OA) and agreement on a positive test (Ps+). Results: The DNA purification methods had very high agreement for categorizing an individual as HPV infected (OA. = 0.95; Ps+. = 0.94) as well as for detection of HPV type-specific infection (OA. = 0.99; Ps+. = 0.88) in ORS. Agreement for detection of HPV type-specific infection was greater than that observed with repeat oral rinse sampling (OA. = 0.99, Ps+. = 0.76) but comparable to inter-assay agreement (OA. = 1.00, Ps+. = 0.90). Conclusions: HPV detection in ORS processed with a magnetic-bead based automated platform will facilitate large natural history studies of oral HPV infection necessary to evaluate the potential use of oral HPV detection in oral cancer screening.
KW - HPV detection
KW - Human papillomavirus
KW - Oral rinse samples
KW - Puregene DNA isolation
KW - Qiasymphony
UR - http://www.scopus.com/inward/record.url?scp=79952627016&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79952627016&partnerID=8YFLogxK
U2 - 10.1016/j.jcv.2010.12.005
DO - 10.1016/j.jcv.2010.12.005
M3 - Article
C2 - 21273118
AN - SCOPUS:79952627016
SN - 1386-6532
VL - 50
SP - 270
EP - 275
JO - Journal of Clinical Virology
JF - Journal of Clinical Virology
IS - 4
ER -