A Comparison of a Prototype PCR Assay and Hybrid Capture 2 for Detection of Carcinogenic Human Papillomavirus DNA in Women With Equivocal or Mildly Abnormal Papanicolaou Smears

Abstract

We evaluated Hybrid Capture 2 (HC2) and polymerase chain reaction (PCR) results for paired specimens collected at 19,187 visits from 5,026 of 5,060 women participating in the Atypical Squamous Cells of Undetermined Significance/Low-Grade Squamous Intraepithelial Lesion Triage Study (ALTS). We examined the test agreement between HC2 and PCR detection for any of 13 carcinogenic human papillomavirus types targeted by HC2 and compared clinical performance of the 2 tests for detecting concurrent and follow-up cervical intraepithelial neoplasia (CIN) 3 or cancer. The k value for the 2 assays was 0.65 (95% confidence interval, 0.64-0.66), with 82.7% crude agreement. HC2 was more sensitive (93.6% vs 89.3%; P < .0005) but less specific (41.2% vs 48.5%; P < .0005) than PCR for detecting 2-year cumulative CIN 3 or cancer (n = 503). The presence of multiple types as detected by PCR and/or cytologic abnormality increased the likelihood of an HC2+ result. Increased sensitivity of HC2 compared with PCR was surprising, given the theoretical advantages of PCR-based methods for analytic sensitivity. Smaller amounts of material used in PCR could have limited its sensitivity, but our results demonstrate the importance of optimization and standardization of PCR-based assays for clinical applications.

Human papillomavirus testing and molecular markers of cervical dysplasia and carcinoma

“…This is an important limitation of the assay, as genotyping can identify the presence of persistent high-risk types that are a risk factor for progression to cervical cancer. 59 Although treatment is currently not defined by specific HPV type, vaccine usefulness could be limited if there is presence of infection by certain types. Advantages of this method include good interlaboratory reproducibility 57 and ease of use.…”

“…This can provide a semiquantitative measure of viral load relative to 1 pg/ml. 57,59,60 However, this assay has no ability to control for the amount of input DNA. The HC2 Assay is formatted to detect only the presence of certain high-risk or low-risk HPV types and will not identify specific types, but it will report that at least 1 of the high-risk or low-risk types is present.…”

Prevent cervical cancer by screening with reliable human papillomavirus detection and genotyping

“…Various HPV DNA tests have been introduced to meet the need for effective cervical cancer prevention screening in China [13,14] and in other countries [15][16][17][18][19][20]. However, these tests without a uniform standard frequently generate discordant results.…”

Performance of the digene LQ, RH and PS HPVs genotyping systems on clinical samples and comparison with HC2 and PCR-based Linear Array

“…This hybridization, strip-based technique allows to identify 37 different HPVs, [19] spanning HR-HPVs, LR-HPVs and other HPVs with still unclear differential association to malignancy. LA has been used in HPVs detection and genotyping in large epidemiological studies [20,21], and has been proposed as a valid methodology for clinical detection of HPVs in paraffin-embedded samples [22,23]. …”