Detection of high-risk human papillomavirus nucleic acid in archival cervical smears.
OBJECTIVE
To test the usefulness of a commercial DNA hybridization assay for the detection of high-risk (HR) human papillomavirus (HPV) types in archival cervical smears and to compare the sensitivity with that of polymerase chain reaction (PCR) using consensus primers.


METHODS
Stained material was scraped from archival slides and the pellet volume noted. DNA was extracted using silica/guanidinium isothiocyanate and the quality checked by amplification of the beta-globin gene. HR-HPV DNA was detected using a commercial hybrid capture assay (HCA) and the results compared with an in-house amplification system with consensus primers.


RESULTS
Of 156 archival smears stored for 12-13 years, 20 were positive by HCA using an HR probe cocktail. Ninety-eight were also tested by PCR, and 35 were positive. The percentage of HPV-positive samples increased with the increasing size of the pellet. HR-HCA detected more positives in samples with high grade squamous intraepithelial lesion (moderate/severe dyskaryosis).


CONCLUSIONS
Both hybridization by HCA and amplification by PCR could be used to detect genital HPV in archival smears. The general primers PCR detected more positives than HR-HCA but included HPV 6/11. While variation in sample size and prolonged storage may reduce the quality of DNA, the use of archival material for longitudinal studies of HPV presence is potentially worthwhile.
Department of Pathology, University of Edinburgh Medical School, U.K.
E
E
McGoogan
A L
AL
Seagar
H A
HA
Cubie
D004279:DNA, Viral / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008137:Longitudinal Studies; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000235:genetics / Q000821:virology*; D010865:Pilot Projects; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D012307:Risk Factors; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000175:diagnosis*; D014626:Vaginal Smears / Q000379:methods
D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic