Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT04167501 |
| Other study ID # |
P/2019/454 |
| Secondary ID |
|
| Status |
Completed |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
February 1, 2020 |
| Est. completion date |
June 1, 2021 |
Study information
| Verified date |
July 2020 |
| Source |
Centre Hospitalier Universitaire de Besancon |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
The objective is to estimate the prevalence of the different genotypes of human
papillomaviruses (HPV) in histologically proven high-grade lesions of the cervix in patients
born between 1972 and 1993. It is plan to determine if the prevalence of HPV16 and HPV18 is
lower in the population born between 1983 and 1993 and potentially exposed to HPV vaccination
compared to those born between 1972 and 1982 who were not exposed to vaccination. Thus the
investigators should be able to determine whether the introduction of HPV vaccination in
France in 2007 has had an impact on the development of high-grade lesions associated with
HPV16 and HPV18.
Description:
The prevalence of the different HPV in high-grade lesions (CIN2/3) will be determined using
the INNOLiPA HPV extra v2 genotyping kit (Fujirebio). Only samples with a histologically
proven high-grade cervical lesion (grade 2 or 3 intraepithelial neoplasia) are concerned. The
patient year of birth, the year of the lesion diagnosis and the histological diagnosis will
be recorded Participating laboratories will select eligible cervical samples (biopsies or
surgical specimen) according to the inclusion criteria defined in the protocol. Only cervical
samples with sufficient biological material will be selected. The selected tissue blocks will
be sent to the Papillomavirus National Reference Center (NRC) by mail (pre-paid envelope
provided) in accordance with the rules for the transport of biological samples. After receipt
by the NRC, the tissue blocks will be anonymized. After HPV genotyping, the blocks will be
returned to the laboratory. Transport costs are covered by the Papillomavirus NRC.
Samples have to be fixed in 4% buffered formaldehyde (maximum 24 hours) and then included in
paraffin for histopathological evaluation. On each of the selected histological pieces, the
papillomavirus NRC will make 5μm thick sections using a microtome. The number of sections to
be made depends on the size of the histological specimen. A minimum surface area equivalent
to 5 sections of 5μm of a 125 mm2 piece is required for genotyping.
Tissue sections are grouped in a labelled 2 mL screw tube. To avoid any contamination between
samples, the microtome blade will be carefully cleaned with xylene and then 95% ethanol
between each block.
After DNA extraction using the QiaAmp DNA mini Kit (Qiagen), HPV genotype analysis will be
performed using molecular techniques for the detection of human papillomavirus DNA. The
investigators will use the INNO-LiPA HPV Genotyping Extra II (Fujirebio) kit. This is a
Polymerase Chain Reaction (PCR) technique using degenerate primers followed by reverse
hybridization on strips on which the specific probes of 32 alpha HPV genotypes are
immobilized. This kit allows the detection of 13 high-risk HPV genotypes (HPV16, 18, 31, 33,
35, 39, 45, 51, 52, 52, 56, 58, 59, 68), 6 HPVs that are likely to be at risk (HPV 26, 53,
66, 70, 73, 82) and 14 low-risk or unclassified HPVs (HPV6, 11, 40, 42, 43, 44, 54, 61, 62,
67, 81, 83, 89). In addition, the samples will be analyzed by PCR luminex according to the
technique published by Schmitt et al, J Clin Microbiol 2010. In addition to alpha HPV, this
technique allows the detection of sixty beta and gamma HPVs.
Characteristics of the subjects to be included A description of patients included in the
study (women born between 1972 and 1982, on the one hand, and women born between 1983 and
1993, on the other) will include age at time of sampling, year of sampling, stage of lesion
CIN2 or 3, and histological type.
Comparisons between age groups will be made using Student tests for quantitative variables
and Chi-square tests for qualitative variables, while respecting the conditions for applying
these tests.
Main judgment criteria Prevalences of high-risk HPV 16 or 18 genotypes will be described in
both age groups with their 95% confidence intervals. These prevalences will be compared
between the two age groups using the Chi-square test.
Secondary Judgment Criterion Prevalences of high-risk HPV genotypes other than 16 or 18 will
be described in both age groups with their 95% confidence intervals. These prevalences will
be compared between the two age groups using the Chi-square test.
Additional analyses The distribution of the different HPV genotypes will be described for
both age groups.
The number of samples required for the study is 606: 303 samples from women born between 1972
and 1982 and 303 samples from women born between 1983 and 1993.
The calculation of the number of samples required is based on the following assumptions: the
prevalence of HPV16 and/or HPV18 in high-grade lesions of women is estimated at 62%. The
minimum decrease after the introduction of HPV vaccination, interesting from a population
health point of view, that the investigators wish to highlight is 10%, i.e. an expected
prevalence among women born between 1983 and 1993 of 52%.
For a power set at 80% and an alpha risk at 5%, a unilateral formulation of hypotheses (only
a decrease in prevalence is possible) and an equal number of samples from women aged 25-34
and 35-44, the number of samples required per group is 303.
