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Type-specific HPV E6/E7 mRNA detection by real-time PCR improves identification of cervical neoplasia.

Journal article
Authors Elin Andersson
Cecilia Kärrberg
Thomas Rådberg
Lennart Blomqvist
Britt-Marie Zetterqvist
Walter Ryd
Magnus Lindh
Peter Horal
Published in Journal of clinical microbiology
Volume 49
Issue 11
Pages 3794-3799
ISSN 1098-660X
Publication year 2011
Published at Institute of Biomedicine, Department of Pathology
Institute of Biomedicine, Department of Infectious Medicine
Institute of Clinical Sciences, Department of Obstetrics and Gynecology
Pages 3794-3799
Language en
Links dx.doi.org/10.1128/JCM.00549-11
Subject categories Cancer and Oncology, Obstetrics and women's diseases

Abstract

DNA-based HPV assays show high sensitivity but poor specificity in detecting high-grade cervical lesions. Assays detecting mRNA of oncogenic E6/E7 show higher specificity, but lack either detection of all high-risk HPV genotypes or the capacity to specify the detected genotypes. Therefore, a real-time PCR assay detecting type-specific E6/E7 mRNA was developed and the clinical performance evaluated. 210 cervical LBC (liquid based cytology) samples from 204 women were analysed for HPV DNA and mRNA with the in house real-time PCR as well as PreTect HPV-Proofer. The sensitivity of real-time PCR mRNA-detection to detect histologically confirmed CIN2+ (cervical intraepithelial neoplasia grade 2 or higher) were 0.91, compared to 0.95 for DNA-analysis. The specificity was 0.68 compared to 0.38, and the positive predictive value (PPV) was higher for mRNA (0.67 vs 0.52) without any loss in negative predictive value (NPV). The sensitivity of the real-time PCR mRNA-test was somewhat higher than for PreTect HPV-Proofer (0.83 vs 0.75), when analysing for the same genotypes. The specificity was similar (0.76 vs 0.77). When analysing for mRNA of the eight most common genotypes in cervical cancer (HPV16, 18, 31, 33, 35, 45, 52, 58), the sensitivity to detect CIN2+ lesions was 0.87 and the specificity 0.74, with a PPV of 0.70. In conclusion, real-time PCR for detection of HPV E6/E7 mRNA transcripts can be a sensitive and specific tool in screening and investigation of cervical neoplasia. The composition of HPV-types in mRNA-testing needs to be further investigated to optimize sensitivity and specificity.

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