Equivocal and weakly positive hybrid capture 2 tests in women aged 50 and older

Abstract

The performance of the Hybrid Capture 2 (HC2) test for human papilloma virus (HPV) detection depends on the prevalence of infection. However, the current HC2 manufacturer recommended interpretative algorithm is the same for all women. This test, which may be particularly useful in perimenopausal and postmenopausal women given the morphologic complexity of their Pap tests, could be affected by the overall lower prevalence of HPV infection in this age group. We investigated HC2 equivocal and weakly positive HPV tests in women 50 years and older and the detection of high-grade dysplasia (CIN2+) on their follow-up specimens. All HC2 test data from 1,067 consecutive specimens and 85 additional specimens from women ≥ 50-years-old with equivocal and weakly positive HC2 were analyzed. Follow-up specimens from women with HC2 tests within these ranges were reviewed. No CIN2+ was found on follow-up of 49 cases of women ≥ 50 with equivocal or weakly positive HC2 results. The current HC2 algorithm resulted in "positive" reports in 63% of specimens with initial equivocal HC2 due to retests mostly within the equivocal range. These results suggest that women 50 years and older may benefit from higher HC2 thresholds. The test could also be reported as HC2 values (RLU/CO) to be interpreted in view of risk factors. Keywordshuman papilloma virus; hybrid capture 2; cervical intraepithelial neoplasia; age Current guidelines recommend cervical cancer screening of perimenopausal and postmenopausal women up to 65-70 years old. 1,2 The interpretation of these Pap tests is challenging because of concurrent hormonally determined cytologic changes. While deep atrophy is characterized by hyperchromasia and increased nuclear cytoplasmic ratios resembling high-grade squamous intraepithelial lesions (HSIL) lesser degrees of atrophy and other perimenopausal changes span the spectrum of reactive changes, atypical squamous cells of undetermined significance (ASCUS) and low grade squamous intraepithelial lesions (LSIL). [3][4][5] Ancillary tests such as high risk human papilloma virus (HPV) detection could therefore be even more important in older women. Using the Hybrid Capture 2 (HC2) system (Qiagen, Valencia, CA) Johnston and Logani showed that HPV testing may be particularly useful in postmenopausal women with ASCUS because a large proportion will be spared from colposcopy given a negative test. 6 However, the performance of HC2 in perimenopausal and postmenopausal women is likely to be affected by a lower prevalence of NIH-PA Author ManuscriptHPV infection 7 as demonstrated in several different populations 8-10 but the current interpretative HC2 algorithm is the same for women of all ages. The HC2 test detects HPV DNA by using probe hybridization and chemiluminescence. The patient's specimen signal as measured in relative light units (RLU) is compared with an average signal of supplied positive reagents. The resulting RLU/cutoff ratio (RLU/CO) is considered positive when ≥ 1 (corresponding to a mean of 1.08 pg/ml of HPV DNA 7 ), but r...

Clinical relevance of the borderline results of the Hybrid Capture 2 High-Risk HPV DNA assay with cervical samples collected in Specimen Transport Medium

“…Ramirez et al 15. and de Vries et al 16. obtained lower values for specimens that retained original HC2 diagnoses after retest (64% and 74%, respectively).…”

“…Since the manufacturer recommends retesting PreservCyt specimens with results within the “grey zone”, several authors have already evaluated whether the retesting algorithm is effective 11, 12, 13, 14, 15, 16. Some authors have performed one retest15, 16, while others have reported two retests13, and some retested only those samples where the results were found within the “grey zone” after the first retest11 or below 1.0 RLU/CO. 14 The range of the “grey zone” varied from 0.8 to 3.0 RLU/CO.…”

“…Several authors9, 10, 11, 12, 13, 14, 15, 16 have investigated the reproducibility and clinical significance of retesting PreservCyt samples with borderline HC2 results. However, we have not found a single study where the same problem was addressed for specimens collected in STM.…”

“Equivocal” High-Risk HPV DNA Tests Performed on ThinPrep Specimens After ASC-US Diagnoses Are Associated With an Increased Incidence of CIN3

“…21,24,28 Regression rates reported in prior studies show that most CIN1 lesion regress without treatment. 21,24,28 Regression rates reported in prior studies show that most CIN1 lesion regress without treatment.…”

Postmenopausal Squamous Atypia: Cytologic Features, Hybrid Capture 2 Tests and Contribution to the ASCUS Pool

“…Albeit in the low range, PSA may on occasion test positive according to the currently used HC2 cutoff of ≥1 RLU/CO [14]. The ideal cutoff value for the HC2 test has been the subject of multiple studies [15][16][17][18][19][20]. For instance, Rebolj et al [15] concluded that HC2 cutoff levels between ≥2 and ≥10 RLU/CO would result in at least 90% sensitivity for CIN-2+ compared to ≥1 RLU/ CO based on their systematic review of the PubMed database.…”