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

Abstract

As part of an ongoing cohort study in the Hokuriku region of Japan, cervical cell samples from histologically confirmed normal (n ‫؍‬ 114) or abnormal (n ‫؍‬ 286) women were examined for the presence of HPV DNA using a second-generation hybrid capture assay (HCA-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 additional 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%. The sensitivity of LCR-E7 PCR increased to 87%, which was significantly higher than that in HCA-II, when testing both highrisk and other HPV types. Sixty-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-II, 5.3% and for LCR-E7 PCR, 1.3%), Key words: hybrid capture assay II; LCR-E7 PCR; human papillomavirus; cervical cancer; high-grade squamous intraepithelial lesionAlthough deaths from cervical cancer in many developed countries have declined in recent decades, it remains the fifth most frequent cancer and the second most common cancer in women worldwide. 1 The adoption of routine cytologic testing for cervical cancer in many developed countries accounts, in large part, for the decrease in deaths from this disease. Problems remain with cytologic testing, however, particularly with the frequency of falsenegatives, the high cost of repeat testing and diagnosis of equivocal cases using the Bethesda system. 2 Given these circumstances, cancer-screening programs using HPV tests are as effective at predicting disease as those using cytologic tests. 3 zur Hausen et al. 4 showed that infection with HPV was closely associated with cervical cancer development. In addition, several previous studies have shown that HPV-6 and -11 are associated with benign anogenital lesions, whereas HPV-16 and -18 are associated with cervical cancer. 5 Currently, more than 80 HPV types have been identified and of these, about 30 distinct HPV types are known to infect the genital tract, 6,7 at least 10 being associated with cancer. 8 Geographic differences in HPV types have been reported to exist between countries 8 and even within the United States. 9,10 The risk factors for cervical cancer and the prevalent HPV types in Japan 11,12 differ from those reported in Western countries. 8 -10 HPV-51, -52 and -58 are more prevalent, whereas HPV-18 and -45 are less prevalent in Japan than in Western countries. 12 Therefore, the definition of high-risk HPV types in Japanese women is important with respect to both clinical management of HPV infection and unequivocal diagnosis of cervical pathology.Highly sensitive HPV DNA tests have been developed as a supplements to cervical cancer screening and fo...

Human papillomavirus infection in Rio de Janeiro, Brazil: a retrospective study

“…High-risk oncogenic HPV types has been found to be the main risk factor for cervical neoplasia [9]; it is also a good predictor of subsequent high-grade lesions in young women [11,12]. Corroborating this proposal, HCA II results were strongly associated with the likelihood of SIL, since we observed that 87% of the patients presenting SIL were also infected by HPV, with a positive trend relating high-risk HPV to cervical transformation (p<0.00001).…”

“…These cases could be latent infections or incipient cervical lesions [13], characterizing a low level of HPV gene replication [10]. However, it also might be due to either DNA testing errors or errors in the cytological diagnosis [9,11]. Viral testing could help to focus on a group of patients with a high risk of developing lesions in the near future, especially in Brazil, due to the high frequency of cervical cancer [15].…”

Detection of human papillomavirus DNA by the hybrid capture assay

“…Other studies have also shown that the amount of HPV DNA is a useful predictor of progression to cervical carcinoma, and concluded that the risk of cancer increased correspondingly [15]. But there have been conflicting reports about correlation between viral load and risk of SIL [16]. Thus, the quantitative aspects of the hybrid capture test merit further evaluation.…”

Detection of human papillomavirus in pterygium and conjunctival papilloma by hybrid capture II and PCR assays

“…One of the advantages of HC-II is its ability to estimate viral genome copy number from the magnitude of the RLU value obtained. 14,15 As the RLU value reflects the activity of HPV, it may be closely associated with the progression of papilloma. 15 In fact, our HC-II data showed that the RLU values in recurrent and re-recurrent papilloma specimens were 25 times and 115 times higher, respectively, than in the primary specimen.…”

“…The second-generation hybrid capture (HC-II) assay is a non-radioactive, reproducible, relatively rapid liquid hybridization assay, with a sensitivity similar to that of PCR. 13,14 It is based on the measurement of relative light units (RLUs) as a semiquantitative index of the amount of HPV DNA. 15 Some previous studies have shown that the RLU level is correlated positively with disease severity.…”