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Journal of Clinical Microbiology, July 2009, p. 2136-2141, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.01733-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

RNA (E6 and E7) Assays versus DNA (E6 and E7) Assays for Risk Evaluation for Women Infected with Human Papillomavirus{triangledown}

Paola Cattani,1* Alessia Siddu,1 Sara D'Onghia,1 Simona Marchetti,1 Rosaria Santangelo,1 Valerio G. Vellone,2 Gian Franco Zannoni,2 and Giovanni Fadda1

Institute of Microbiology,1 Institute of Pathology, Università Cattolica del Sacro Cuore, Largo A. Gemelli 8, 00168 Rome, Italy2

Received 8 September 2008/ Returned for modification 1 March 2009/ Accepted 21 April 2009

In the majority of cases, high-risk human papillomavirus (HR HPV) infections regress spontaneously, with only a small percentage progressing to high-grade lesions. Current screening methods are based on DNA detection. An alternative would be to monitor expression of the E6 and E7 viral oncogenes continuously expressed by malignant phenotypes. In the work reported in this paper, we compared the two methods for a group of women with high-risk HPV infections. Cervical specimens from 400 women, previously found to be HPV DNA positive, were analyzed for HPV DNA by a liquid hybridization assay and typed by multiplex PCR (for types 16, 18, 31, and 33). Identification of HR HPV E6 and E7 RNA transcripts was performed using real-time reverse transcription-PCR and nucleic acid sequence-based amplification assays. Results were compared with concurrent cytological data. HR HPVs were found in 61.2% of patients. The most common genotype was HPV type 16 (HPV-16) (47.1%), followed by HPV-18, HPV-31, and HPV-33. Nine percent of cases involved other genotypes. Among 223 HPV DNA-positive samples, only 118 were positive in the RNA test. Among HPV DNA-positive patients with normal cytology, we detected E6 and E7 RNA transcripts in two cases (18.2%). The rate of detection increased gradually with the grade of the observed lesions, rising from 20% for patients with atypical squamous cells of undetermined significance to 48.1% for women with low-grade squamous intraepithelial lesions and 86.3% for those with high-grade squamous intraepithelial lesions. These results suggest that testing for HPV E6 and E7 transcripts could be a useful tool for screening and patient management, providing more accurate predictions of risk than those obtained by DNA testing.

* Corresponding author. Mailing address: Institute of Microbiology, Università Cattolica del Sacro Cuore, Largo A. Gemelli 8, 00168 Rome, Italy. Phone: 39 06 30154964. Fax: 39 06 3051152. E-mail: pcattani{at}rm.unicatt.it

{triangledown} Published ahead of print on 29 April 2009.

Journal of Clinical Microbiology, July 2009, p. 2136-2141, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.01733-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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