Hybrid capture 2 test results after an initial equivocal RLU/CO value are dependent on age

The effect of age on Hybrid Capture 2 (HC2) tests initially falling within the equivocal range has not been determined. We identified 359 cervicovaginal liquid cytology specimens with initial equivocal values. First and second retest relative light units/cutoff (RLU/CO) values were compared for women of 3 different age groups (15-29, 30-49, and ≥50 years). The proportion of first retests with an RLU/CO of less than 1 increased with age (P < .001). Of the 56 second retests performed, only 4 had an RLU/CO of 1 or more. The proportion of "positive" HC2 results following the current HC2 algorithm decreased with increasing age (P < .001), showing that HC2 test results after an initial equivocal value are dependent on age. Follow-up demonstrated cervical intraepithelial neoplasia grade 2 or worse (CIN2+) in 6 (5.9%) women 15 to 29 years old and in 5 (6.3%) women 30 to 49 years old. No CIN2+ was found on follow-up of 34 of 57 women 50 years and older. These results likely reflect human papillomavirus infection prevalence and question the use of identical cutoff values regardless of age for detection of CIN2+.