Human papillomavirus load in Hybrid Capture II assay: does increasing the cutoff improve the test?
OBJECTIVE
The aim of the study was to evaluate whether the results of the Hybrid Capture II (HCII) assay for detecting high-grade squamous intraepithelial lesions (H-SIL) or cervical carcinoma can be improved by increasing the relative light units (RLU) level.


METHODS
We included 2271 women (mean age 38.7 +/- 12.3, range 15-92) referred to a colposcopic clinic due to cytology of atypical cells of unknown significance, SIL or carcinoma. All women underwent a new Pap test, HR-HPV detection using HCII and colposcopy with biopsy of suspicious areas when present.


RESULTS
HR-HPV was detected in 91.7% of carcinomas, 96.6% of H-SIL, 85.1% of low-grade SIL and 21.6% of cases with no lesion. The probability of harboring an H-SIL or a carcinoma significantly increased as RLU increased (P = 0.01). The sensitivity and specificity for H-SIL or carcinoma at different cutoffs were 95.7 and 54.6 at 1 RLU, 93.9 and 59.6 at 2 RLU, 90.1 and 65.1 at 5 RLU and 85.7 and 68.7 at 10 RLU. The percentage of cases not detected with HCII increased from 2.4% for cases with <1 RLU to 9.5% for cases between 1 and 2 RLU, 14.8% between 2 and 3 RLU, 21.7% between 3 and 5 RLU and 28.4% between 5 and 10 RLU.


CONCLUSIONS
The use of a higher cutoff (higher viral load) in the HCII should not be recommended because it significantly increases the number of cases with H-SIL or carcinoma not detected, reducing the sensitivity and negative predictive value of the test.
Ordi
Jaume
J
Department of Anatomical Pathology, IDIBAPS-Hospital Clínic, University of Barcelona, School of Medicine, C/Villarroel 170, 08036-Barcelona, Spain. jordi@clinic.ub.es
Alonso
Imma
I
Torné
Aureli
A
Esteve
Roser
R
Sierra
Erika
E
Campo
Elias
E
Puig-Tintoré
Luis M
LM
D000230:Adenocarcinoma / Q000175:diagnosis / Q000473:pathology / Q000821:virology; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000473:pathology / Q000821:virology; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D006801:Humans; D008875:Middle Aged; D009693:Nucleic Acid Hybridization / Q000379:methods; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000150:complications / Q000175:diagnosis* / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000473:pathology / Q000821:virology; D019562:Viral Load; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis* / Q000473:pathology / Q000821:virology
D004279:DNA, Viral; D012367:RNA, Viral