Hybrid capture-II and LCR-E7 PCR assays for HPV typing in cervical cytologic samples

As part of an ongoing cohort study in the Hokuriku region of Japan, cervica l cell samples from histologically confirmed, or abnormal (n=286) women wer e examined for the presence of HPV DNA using a second-generation hybrid cap ture assay (HICA-II) and LCR-E7 PCR. HCA-II detected low-risk (HPV-6, -11, -42, 43 and -44) and high-risk (HPV-16, -18, -31, -33, -35, -39, -45, -51, -52, -56, -58, -59 and -68) HPV types, while LCR-E7 PCR detected an additio nal 7 HPV types and some uncharacterized types. In screening of high-grade squamous intraepithelial lesions (HSILs) and invasive cervical cancer, the sensitivities of HCA-II and LCR-E7 PCR testing the high-risk HPV types were 83% and 81%, respectively, while the specificity of both assays was 93%. T he sensitivity of LCR-E7 PCR increased to 87%, which was significantly high er than that in HCA-II, when testing both highrisk and other HPV types. Six ty-eight inconsistent results (17% of total tested) from HCA-II and LCR-E7 PCR were due to (I) low copy number of HPV genome (false-negative for HCA-I I, 5.3% and for LCR-E7 PCR, 1.3%), (ii) infection with HPV types undetectab le by HCA-II (4.8%), (iii) multiple HPV infections (5%) or (iv) unknown rea sons (0.8%). LCR-E7 PCR revealed that infections with HPV-16, -18, -31, -33 , -35, -51, -52, -56, -58 or -67 was a high risk for cancer since these typ es predominated in HSIL and invasive cervical cancer. Samples showing high relative light units (>20) with a high-risk probe in HCA-II also gave posit ive results in LCR-E7 PCR and were generally associated with abnormal cervi cal lesions. Thus, we propose that both HCA-II and LCR-E7 PCR are valuable screening tests for premalignant and malignant cervical lesions. (C) 2001 W iley-Liss, Inc.