Optimal Threshold for a Positive Hybrid Capture 2 Test for Detection of Human Papillomavirus: Data from the ARTISTIC Trial

Abstract

We present data on the use of the Hybrid Capture 2 (HC2) test for the detection of high-risk human papillomavirus (HR HPV) with different thresholds for positivity within a primary screening setting and as a method of triage for low-grade cytology. In the ARTISTIC population-based trial, 18,386 women were screened by cytology and for HPV. Cervical intraepithelial neoplasia lesions of grade two and higher (CIN2؉ lesions) were identified for 453 women within 30 months of an abnormal baseline sample. When a relative light unit/cutoff (RLU/Co) ratio of >1 was used as the threshold for considering an HC2 result positive, 15.6% of results were positive, and the proportion of CIN2؉ lesions in this group was 14.7%. The relative sensitivity for CIN2؉ lesion detection was 93.4%. When an RLU/Co ratio of >2 was used as the threshold, there was a 2.5% reduction in positivity, with an increase in the proportion of CIN2؉ lesions detected. The relative sensitivity decreased slightly, to 90.3%. Among women with low-grade cytology, HPV prevalences were 43.7% and 40.3% at RLU/Co ratios of >1 and >2, respectively. The proportions of CIN2؉ lesions detected were 17.3% and 18.0%, with relative sensitivities of 87.7% at an RLU/Co ratio of >1 and 84.2% at an RLU/Co ratio of >2. At an RLU/Co ratio of >1, 68.3% of HC2-positive results were confirmed by the Roche line blot assay, compared to 77.2% of those at an RLU/Co ratio of >2. Fewer HC2-positive results were confirmed for 35-to 64-year-olds (50.3% at an RLU/Co ratio of >1 and 63.2% at an RLU/Co ratio of >2) than for 20-to 34-year-olds (78.7% at an RLU/Co ratio of >1 and 83.7% at an RLU/Co ratio of >2). If the HC2 test is used for routine screening as an initial test or as a method of triage for low-grade cytology, we would suggest increasing the threshold for positivity from the RLU/Co ratio of >1, recommended by the manufacturer, to an RLU/Co ratio of >2, since this study has shown that a beneficial balance between relative sensitivity and the proportion of CIN2؉ lesions detected is achieved at this threshold.Persistent infection with any of the 15 cancer-associated high-risk human papillomavirus (HR HPV) genotypes is now well recognized as essential for the subsequent development of cervical cancer and its high-grade precursor lesions (2,15,16). Due to the very high prevalence of HPV infections that typically resolve within 1 to 2 years, especially in younger women (6, 11), the role of HPV testing in the early detection of cervical lesions remains controversial. The most widely used test for the detection of a group of 13 HR HPV genotypes is the commercially available, FDA-approved Hybrid Capture 2 high-risk HPV DNA test (HC2 test; Qiagen [formally known as Digene]). Hybrid Capture 2 technology consists of a nucleic acid hybridization assay with signal amplification that utilizes microplate chemiluminescence for the qualitative detection of HPV. There is increasing interest in the use of HC2 technology within the cervical screening program either as a stand-alone screening test or i...

Correlation between hybrid capture II high‐risk human papillomavirus DNA test chemiluminescence intensity from cervical samples with follow‐up histologic results

“…[24][25][26][27][28][29] The recently published series from Sargent et al 29 reported a reduced rate of high-grade dysplasia after an hrHPV test, with an RLU/CO value of between 1 and 2 (ie, 9.2% vs 17.9% for an RLU/CO value 2.0), although their methodology differs considerably from that of the current present study (eg, combining ''borderline'' and ''mild'' cytology). Nevertheless, the indicated rate of 9.2% represents a nearly 5-fold risk compared with that associated with negative hrHPV tests in the study by Sargent et al 29 Using polymerase chain reaction-based hrHPV detection, an increase in the viral load of carcinogenic subtypes demonstrates a positive correlation with cervical abnormalities in the general population (ie, when all Pap diagnoses are included), 26 and the association between type-specific HPV-16 viral load with CIN III or cancer indicates a positive correlation in patients with negative cervical cytology, but not those with ASC-US cytology. 24 When HC II was used to detect hrHPV subtypes, no association was noted between increasing viral load and the risk of CIN III or cancer, 27 although a modest positive correlation was observed for CIN I.…”

Semi‐quantitative HPV viral load in patients with ASC‐US cytology: viral load correlates strongly with the presence of CIN but only weakly with its severity

“…To improve the test specificity, several studies have implemented a higher cut-off for the positivity of HPV DNA testing. 8 However, these trials caused a decrease in sensitivity and negative predictive value (NPV), and no cut-off level could exclude patients with underlying CIN2/CIN3 lesions or cancer. However, the use of viral load as a predictor of high-grade cervical lesions in women with ASC-US cytology is doubtful, because of the large overlap of diagnosis.…”

Human papillomavirus viral load in cervical intraepithelial neoplasia as a prognostic factor in a Mexican population

“…On the spectrum of cervical disease, CIN2 and 3 are considered cervical cancer precursors. Several studies have consistently reported that the persistence of HR-HPV in the genital tract is necessary for the development and progression of cervical dysplastic lesions (Ho et al, 1995;Remmink et al, 1995;Nobbenhuis et al, 1999;Briolat et al, 2007;Xi et al, 2008;Sargent et al, 2010). Conversely, high-grade squamous intraepithelial lesions were shown to be unlikely to develop in HR-HPVnegative women in a 2-year follow-up cohort, and smears showed that mild or borderline cell atypia returns to normal (Nobbenhuis et al, 2001;Zielinski et al, 2001).…”

“…Specimens with a ratio of <1.00 were considered to be negative, and those with a ratio of ≥1.00 were considered to be positive because they contained at least 1 pg/mL HPV DNA, equivalent to 100,000 HPV copies/mL or 5000 HPV copies/assay (Sargent et al, 2010).…”