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Viral DNA load, physical status and E2/E6 ratio as markers to grade HPV16 positive women for high-grade cervical lesions.
Gynecol Oncol. 2007 Sep; 106(3):549-57.GO

Abstract

OBJECTIVES

Cervical intraepithelial neoplasias (CIN) associated with high-risk (HR) human papillomavirus infection, in addition to HR-HPV typing need other viral marker testing to distinguish a subset of lesions with clinical relevant infections. This study has evaluated the significance of viral markers, such as viral load, physical status and E2/E6 ratio, to stratify HPV16 infected women at a single point in time for grade of cervical lesions.

METHODS

One hundred sixty-six cytological specimens were selected from women with low (n=72) and high (n=94) grade squamous intraepithelial lesions (SIL), and positive to HPV16. All the 72 LSIL were CINI, 83 of the 94 HSIL were CINII/III and 11 SCC (Squamous Cervical Carcinoma). Cytological specimens were analysed by two different SYBR Green Real-time PCR assays (RT-PCR). Specific primers for both E2 and E6 viral genes and GAPDH cellular gene were designed to determine viral load, physical status and E2/E6 ratio.

RESULTS

The viral load was significantly higher in HSIL than in LSIL. In CINI episomal DNA was prevalent (72.2%), mixed forms (episomal and integrated) were 27.8%, suggestive of an early integration of viral DNA into cellular genome, no pure integrated forms were detected. However in CINII/III mixed DNA forms were prevalent (73.5%). In SCC pure integrated DNA was prevalent (81.8%) in absence of episomal forms. E2/E6 ratio decreased significantly from CINI to CINII/III and SCC with a linear trend. The logistic regression analysis showed that viral load higher than 1.38x10(6) genome copies per 300 ng of total DNA associated with E2/E6 ratio lower than 0.90 was highly significant in differentiating CINII/III versus CINI, while the only E2/E6 value lower than 0.17 was significant in differentiating SCC from CINI.

CONCLUSIONS

Viral load higher than 1.38x10(6) genome copies per 300 ng of total DNA and E2/E6 ratio values allow HPV16 infected women with high grade cervical intraepithelial lesions to be recognized.

Authors+Show Affiliations

Department of Clinical and Experimental Medicine, Microbiology Section, University of Bologna, and Department of Obstetrics and Gynecology, S. Orsola-Malpighi Hospital, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17568661

Citation

Cricca, Monica, et al. "Viral DNA Load, Physical Status and E2/E6 Ratio as Markers to Grade HPV16 Positive Women for High-grade Cervical Lesions." Gynecologic Oncology, vol. 106, no. 3, 2007, pp. 549-57.
Cricca M, Morselli-Labate AM, Venturoli S, et al. Viral DNA load, physical status and E2/E6 ratio as markers to grade HPV16 positive women for high-grade cervical lesions. Gynecol Oncol. 2007;106(3):549-57.
Cricca, M., Morselli-Labate, A. M., Venturoli, S., Ambretti, S., Gentilomi, G. A., Gallinella, G., Costa, S., Musiani, M., & Zerbini, M. (2007). Viral DNA load, physical status and E2/E6 ratio as markers to grade HPV16 positive women for high-grade cervical lesions. Gynecologic Oncology, 106(3), 549-57.
Cricca M, et al. Viral DNA Load, Physical Status and E2/E6 Ratio as Markers to Grade HPV16 Positive Women for High-grade Cervical Lesions. Gynecol Oncol. 2007;106(3):549-57. PubMed PMID: 17568661.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Viral DNA load, physical status and E2/E6 ratio as markers to grade HPV16 positive women for high-grade cervical lesions. AU - Cricca,Monica, AU - Morselli-Labate,Antonio Maria, AU - Venturoli,Simona, AU - Ambretti,Simone, AU - Gentilomi,Giovanna Angela, AU - Gallinella,Giorgio, AU - Costa,Silvano, AU - Musiani,Monica, AU - Zerbini,Marialuisa, Y1 - 2007/06/13/ PY - 2007/02/01/received PY - 2007/04/27/revised PY - 2007/05/07/accepted PY - 2007/6/15/pubmed PY - 2007/10/12/medline PY - 2007/6/15/entrez SP - 549 EP - 57 JF - Gynecologic oncology JO - Gynecol. Oncol. VL - 106 IS - 3 N2 - OBJECTIVES: Cervical intraepithelial neoplasias (CIN) associated with high-risk (HR) human papillomavirus infection, in addition to HR-HPV typing need other viral marker testing to distinguish a subset of lesions with clinical relevant infections. This study has evaluated the significance of viral markers, such as viral load, physical status and E2/E6 ratio, to stratify HPV16 infected women at a single point in time for grade of cervical lesions. METHODS: One hundred sixty-six cytological specimens were selected from women with low (n=72) and high (n=94) grade squamous intraepithelial lesions (SIL), and positive to HPV16. All the 72 LSIL were CINI, 83 of the 94 HSIL were CINII/III and 11 SCC (Squamous Cervical Carcinoma). Cytological specimens were analysed by two different SYBR Green Real-time PCR assays (RT-PCR). Specific primers for both E2 and E6 viral genes and GAPDH cellular gene were designed to determine viral load, physical status and E2/E6 ratio. RESULTS: The viral load was significantly higher in HSIL than in LSIL. In CINI episomal DNA was prevalent (72.2%), mixed forms (episomal and integrated) were 27.8%, suggestive of an early integration of viral DNA into cellular genome, no pure integrated forms were detected. However in CINII/III mixed DNA forms were prevalent (73.5%). In SCC pure integrated DNA was prevalent (81.8%) in absence of episomal forms. E2/E6 ratio decreased significantly from CINI to CINII/III and SCC with a linear trend. The logistic regression analysis showed that viral load higher than 1.38x10(6) genome copies per 300 ng of total DNA associated with E2/E6 ratio lower than 0.90 was highly significant in differentiating CINII/III versus CINI, while the only E2/E6 value lower than 0.17 was significant in differentiating SCC from CINI. CONCLUSIONS: Viral load higher than 1.38x10(6) genome copies per 300 ng of total DNA and E2/E6 ratio values allow HPV16 infected women with high grade cervical intraepithelial lesions to be recognized. SN - 0090-8258 UR - https://www.unboundmedicine.com/medline/citation/17568661/Viral_DNA_load_physical_status_and_E2/E6_ratio_as_markers_to_grade_HPV16_positive_women_for_high_grade_cervical_lesions_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0090-8258(07)00339-3 DB - PRIME DP - Unbound Medicine ER -