Evaluation of a Newly Developed GenoArray Human Papillomavirus (HPV) Genotyping Assay and Comparison with the Roche Linear Array HPV Genotyping Assay

Persistent infection with high-risk types of human papillomavirus (HPV) is a necessary step in the development of cervical cancer. The incorporation of HPV detection into cervical screening programs may improve the ability to identify women at risk of cervical cancer. We recently evaluated the performance characteristics of a newly developed HPV detection assay, the GenoArray (GA) genotyping assay, for the detection of HPV infections by comparing it with the commercial Roche Linear Array (LA) HPV genotyping assay. The GA assay has an analytical sensitivity for the detection of HPV types 16 (HPV-16) and HPV-18 of as few as 10 to 50 copies, and its reproducibility is adequate. The GA and LA assays showed no significant difference in the rates of detection of genotypes detected by both HPV genotyping assays and oncogenic genotypes, and the interassay agreement was excellent. The GA and LA assays revealed either concordant or compatible genotyping results for 97.5% of the samples and discordant results for only eight (2.5%) samples. Compatible results were also observed for the detection of single or multiple HPV infections and the detection of most of the genotypes. The GA assay also demonstrated good clinical performance characteristics when the comparisons were carried out with clinical subgroups of samples from patients with normal cytologies, low-grade or high-grade squamous intraepithelial lesions, and cancers. Therefore, the GA assay appears to be highly sensitive and specific for the genotyping of HPV. It has the advantage that it specifically detects HPV-52, which overcomes a limitation of the LA assay, and hence, it has potential value for use for genotyping, especially in regions where HPV-52 has a high prevalence.Human papillomavirus (HPV) is the main etiological agent for the development of cervical cancer. Persistent infection with high-risk (HR) HPV types is recognized as a necessary step in the progression to neoplastic disease (26). Tests for HPV DNA have shown encouraging results when they are used in conjunction with the traditional cytology screening method (3, 7, 8). Many molecular methods for testing for HPV are currently available. The Digene/Hybrid Capture II (HC II) commercial HPV detection kit and the Cervista HPV HR and Cervista 16/18 tests are approved by the FDA for use for routine screening for HPV (9, 19). However, both the HC II and the Cervista HPV HR assays are unable to discriminate specific genotypes or to identify infections involving multiple genotypes, and the Cervista assay detects only two HPV types (types 16 and 18). Recent studies have suggested that different high-risk HPV types have different oncogenic potentials (5). Besides the clinical aspect, HPV genotyping is also a key part of epidemiological studies of HPV infections worldwide.A number of PCR-based HPV genotyping assays have been developed to amplify HPV DNA, followed by reverse hybridization against immobilized genotype-specific probes, allowing the simultaneous identification of a broad range of anogenital HPV genotypes. However, the different assays have potential variations in their abilities to detect different HPV types due to their different analytical sensitivities and specificities and their failure to detect specific variants (2, 17, 20).The HPV GenoArray (GA) test (Hybribio Ltd., Hong Kong) is a newly developed PCR-based HPV genotyping assay. It utilizes L1 consensus primers to simultaneously amplify 21 HPV genotypes, followed by flowthrough hybridization with immobilized genotype-specific probes. It is marked as Conformité Européenne (CE) for use in Europe and is currently being used in some hospitals in China.The present study was designed to evaluate the analytical and clinical performance characteristics of the Hybribio GA test, and the level of concordance of the genotypes detected by the GA test was compared to that of the Roche Linear Array (LA) HPV test. We focused on interassay agreement for overall HPV DNA detection and for the detection of HPV oncogenic risk types. We also assessed the interassay agreement for type-specific as well as single and multiple HPV infections.