A Comparison of Linear Array and Line Blot Assay for Detection of HPV and Cervical Precancer and Cancer in the ASCUS LSIL Triage Study

Division of Cancer Epidemiology and Genetics and Division of Cancer Prevention, National Cancer Institute, NIH, DHHS, Bethesda, MD 20892, USA; Departments of Epidemiology and Molecular Microbiology and Immunology, Johns Hopkins University, Baltimore, MD 21205, USA; Departments of Molecular Genetics and Microbiology and Obstetrics and Gynecology, University of New Mexico Health Sciences Center, School of Medicine, Albuquerque, NM, 87131, USA

^* To whom correspondence should be addressed. Email: castlep{at}mail.nih.gov.

	Abstract

We evaluated Linear Array (LA), a newly commercialized PGMY09/11 L1 consensus primer PCR test that detects 37 human papillomavirus (HPV) genotypes by reverse line blot hybridization, for the detection of individual HPV genotypes and carcinogenic HPV and its clinical performance for detecting 2-year cumulative cervical precancer and cancer using archived specimens from the ASCUS and LSIL Triage Study (ALTS). LA testing was conducted on enrollment specimens from women referred because of an ASCUS Pap. To gauge the performance of the new test, results were compared to its prototype predecessor assay, Line Blot Assay (LBA), restricted to paired results (n = 3,335). LA testing was done masked to LBA results and clinical outcomes. The results of LA and LBA testing were compared for detection of carcinogenic HPV and clinical outcomes of cervical precancer and cancer. Overall, 50% and 55% of women tested positive for carcinogenic HPV by LBA and LA, respectively (P < 0.0001). The percent agreement for carcinogenic HPV detection was 88%, percent positive agreement was 80%, and kappa was 0.76 for detection of carcinogenic HPV by the two assays. There was a significant increase in detection by LA for most of the 37 HPV genotypes targeted by both assays, including for 13 of 14 carcinogenic HPV genotypes. LA detected more multiple-genotype infections for all HPV genotypes among HPV-positive women (P < 0.0001) and for carcinogenic HPV genotypes among carcinogenic HPV-positive women (P < 0.0001). LA was more sensitive (92.3% vs. 87.1%; P = 0.003) and less specific (48.2% vs. 54.0%; P < 0.0001) than LBA for 2-year cumulative cervical precancer and cancer as diagnosed by the quality control pathology group. In conclusion, we found LA to be a promising assay for the detection of HPV genotypes and carcinogenic HPV, and may be clinically useful for the detection of cervical precancer and cancer in women with equivocal cytology.