OC 05-01HIGH-RISK HPV DETECTION IN PLASMA SAMPLES OF WOMEN WITH A RECENT HISTORY OF CERVICAL DYSPLASIA.

02. Epidemiology and natural history
M. Martinelli 1, R. Musumeci 1, E. Calaresu 1, E. Tanzi 2, G. Sotgiu 3, A. Piana 3, C. Cocuzza 1.
1Department of Medicine and Surgery, University of Milano-Bicocca (Italy), 2Department of Biomedical Sciences for Health, University of Milano (Italy), 3Department of Biomedical Sciences, University of Sassari (Italy)

Background / Objectives

Circulating HPV DNA has been previously described in women with advanced stages of cervical cancer and has been suggested to be a prognostic marker of disease recurrences and metastases1,2. The presence of HPV in plasma of women with early stages of cervical dysplasia has not been previously reported. The aim of this study was to investigate the presence of high-risk HPV (hrHPV) in cervical and plasma samples of women with a recent history of cervical dysplasia.

 


Methods

Blood and cervical samples were obtained from 120 women referred to San Gerardo Hospital, Monza, Italy with a recent history of ASCUS or low-grade cervical dysplasia (L-SIL). Routine  cervical cytology (Pap test) and hrHPV detection from cervical and plasma samples were performed on all samples. A control group of 20 healthy females was included for hrHPV testing in plasma. Nucleic acid extraction was performed using automated NucliSENS easyMAG system (bioMérieux). HPV 16, 18, 31, 33, 45, 51 and 52 detection was carried out by means of previously described genotype-specific "in house" real-time PCR assays3, 4, 5 and confirmed by Anyplex II HPV28 (Seegene); all assays were validated through the participation the Laboratory to the WHO LabNet HPV Proficiency Study.


Results

At the time of blood collection, cervical cytology results showed 26 women to have regressed to a negative Pap test, 24 with ASCUS, 52 L-SIL and 18 H-SIL.

Overall hrHPV positivity for one or more of the tested high-risk genotypes showed rates of 44.2% (53/120) in cervical samples, 34.2% (41/120) in plasma, and 20.8% (25/120) of women were found to have hrHPV in both blood and cervix.

In particular, plasma HPV DNA positivity rates according to cervical cytology results at the time of blood sample collection were: negative cytology 30.8% (8/26); ASCUS 33.3% (8/24); L-SIL 30.7% (16/52); H-SIL 50% (9/18). No HPV positivity was found in blood samples of women from the control group.

HPV 45 (46.2%), 51 (30.8%) and 16 (17.3%) were found to be the most prevalent genotypes in positive plasma samples whilst HPV 31 and 33 were not detected.


Conclusion

In this study, real-time PCR detected HPV 45, 51, 16, 52 and 18 DNA in the peripheral blood of women with a recent history of ASCUS or low-grade cervical dysplasia, with positivity rates ranging from 30.7% to 50% depending on cervical cytology results at the time of blood collection. This is to our knowledge the first report of hrHPV DNA detection in plasma samples of women with an early history of cervical dysplasia. Further studies are required to evaluate the significance of hrHPV DNA detection in the circulatory system of women with transient or early stages of cervical dysplasia.


References

1. Sathish N et al. J Clin Virol 2004; 31:204-9

2. Wei Y C et al. Int J Gynaecol Obstet 2007; 96:112-6

3. Broccolo F, Cocuzza C. J Med Virol 2009; 81:278-287

4. Lindh M et al. J Clin Virol 2007; 40:321-324

5. Flores-Munguia R et al. J Mol Diagn 2004; 6; 2:115-124