Comparison of the Abbott RealTi m e High-Risk Human Papillomavirus (HPV), Roche Cobas HPV, and Hybrid Capture 2 Assays to Direct Sequencing and Genotyping of HPV DNA

Abstract

bInfection with high-risk (HR) human papillomavirus (HPV) genotypes is an important risk factor for cervical cancers. We evaluated the clinical performances of two new real-time PCR assays for detecting HR HPVs compared to that of the Hybrid Capture 2 test (HC2). A total of 356 cervical swab specimens, which had been examined for cervical cytology, were assayed by Abbott RealTime HR and Roche Cobas HPV as well as HC2. Sensitivities and specificities of these assays were determined based on the criteria that concordant results among the three assays were regarded as true-positive or -negative and that the results of genotyping and sequencing were considered true findings when the HPV assays presented discrepant results. The overall concordance rate among the results for the three assays was 82.6%, and RealTime HR and Cobas HPV assays agreed with HC2 in 86.1% and 89.9% of cases, respectively. The two real-time PCR assays agreed with each other for 89.6% of the samples, and the concordance rate between them was equal to or greater than 98.0% for detecting HPV type 16 or 18. HC2 demonstrated a sensitivity of 96.6% with a specificity of 89.1% for detecting HR HPVs, while RealTime HR presented a sensitivity of 78.3% with a specificity of 99.2%. The sensitivity and specificity of Cobas HPV for detecting HR HPVs were 91.7% and 97.0%. The new real-time PCR assays exhibited lower sensitivities for detecting HR HPVs than that of HC2. Nevertheless, the newly introduced assays have an advantage of simultaneously identifying HPV types 16 and 18 from clinical samples. P ersistent cervical infection of human papillomavirus (HPV) is a well-known risk factor for developing cervical cancer, which is the second most common malignancy in women, causing approximately 250,000 deaths each year worldwide (35, 36). More than 150 HPV genotypes have been identified, and approximately 50 of them are known to infect cervical epithelia. Recently, the International Agency for Research on Cancer (IARC) defined 12 high-risk (HR) HPV genotypes (HPV types 16,18,31,33,35, 39, 45, 51, 52, 56, 58, and 59) as group 1 carcinogens (3). In particular, HPV16 is known as the most common HPV type, contributing to approximately half of all invasive cervical cancer cases. This genotype is also known to demonstrate a biological advantage in transmission, persistence, and transformation (2). HPV18 has been reported as the second most common cause of HPV-associated invasive cervical cancers, and the proportion of cervical cancers related to HPV16 and/or HPV18 has been reported to be between 68% and 82% (19).Morphological examination of cervix cytology has been used as a screening test for the prevention and early detection of cervical cancers over the past 50 years (12, 13). A major target of the cervical screening is to detect cases of cervical intraepithelial neoplasia grade 3 (CIN3), which can be treated before it progresses to invasive cancer. However, cytology tests are limited in predicting HPV infections and cervical cancers. Abnormal findings in cervic...

Testing and Genotyping of High-Risk Human Papillomavirus by the cobas HPV Test and the Hybrid Capture 2 High-Risk HPV DNA Test Using Cervical and Vaginal Samples

“…Cross-reactions with nononcogenic HPV genotypes (10,11) have also been shown for the hc2 assay. Contrary to our results, Park et al (12) showed a higher proportion of samples that were positive by the hc2 test and negative by the cobas test, as well as a higher positivity rate for the hc2 test than the cobas test; Wong et al (13) showed a higher confirmation rate by the Linear Array test for the hc2 test than for the cobas test.…”

A Comparison of the Roche Cobas HPV Test With the Hybrid Capture 2 Test for the Detection of High-Risk Human Papillomavirus Genotypes

“…This is in contrast to a previous study, which compared the HC2 and cobas assay using ThinPrep preparation and reported a higher sensitivity with the HC2 assay. 12 One difference in the prior study is that all specimens were collected in PreservCyt transport medium, whereas only a minority of cases were collected in PreservCyt solution in the present study. Another notable difference between the previous and our current study is that the specimens were stored at À708C for unknown durations before being tested in the former study, whereas the specimens in the present study were stored at room temperature and were analyzed within 1 week after collection.…”

“…20 Based on the ATHENA study, a slightly higher positive hrHPV rate was also noted with the cobas assay compared with the HC2 assay (32.6% versus 31.5%) among women with atypical squamous cells of undetermined significance, 14 whereas other studies observed a higher positive hrHPV rate with the HC2 assay compared with the cobas assay in specimens collected with Preservcyt Solution (Hologic). [10][11][12] Approximately 80% (7 of 9) of HC2-positive/cobasnegative cases tested positive for low-risk HPV only by LA whereas 13.3% (4 of 30) of cobas-positive/HC2-negative cases did. This difference was statistically significant.…”

“…The cobas test has been validated in several studies using either ThinPrep LB Pap specimens or specimens collected in specimen transport medium (Qiagen), but neither cobas nor HC2 has been approved for use with SurePath LB Pap specimens (BD Diagnostic, Burlington, Massachusetts). [9][10][11][12][13][14] As part of the validation process, we compared the performance of the cobas HPV test with our in-house validated HC2 assay for the detection of hrHPV DNA using both SurePath and ThinPrep preparations.…”

Look for Efficiency: Real Time PCR Using Cobas 4800 - An Efficient Tool for HPV Screening

“…They found a strong agreement between the cobas 4800 HPV test and HC2 (97.3% and 98.3% for controls and cases, respectively. Similarly, Park et al, [29] demonstrated, with HC2, a sensitivity of 96.6% with a specificity of 89.1% for detecting HR HPVs, while, with Cobas, they were 91.7% and 97.0%, respectively, thus very close. These 2 studies involved a relatively small sample (less than 1000 and 356 respectively).…”