This prospective, descriptive, analytical study included 27 women attending the gynecology department of the Thies Regional Hospital Centre (CHRT), Senegal, for cervical cancer screening or follow-up of precancerous lesions. Written informed consent was obtained from all participants.

Cervical-vaginal samples were collected using sterile cytobrushes to recover both endocervical and exocervical cells. Prior to sampling, participants were checked for the absence of interfering substances (creams, gels, vaginal douches). Excess mucus was removed, and the cytobrush was inserted 1 - 1.5 cm into the cervix and rotated three times. Samples were placed in PreservCyt^® tubes, transported at 2˚C - 30˚C, and stored at −20˚C until DNA extraction.

DNA was extracted using the Quick-DNA^TM Miniprep Plus kit (Zymo Research) according to the manufacturer’s instructions, with sterile RNase-free water as extraction control. Briefly, 200 µL of sample were lysed with 200 µL BioFluid & Cell buffer and 20 µL Proteinase K, vortexed, and incubated at 55˚C for 10 min. Lysates were processed through Zymo-Spin^TM IIC-XLR columns, washed sequentially with DNA Pre-Wash and g-DNA Wash buffers, and DNA was eluted with ≥50 µL elution buffer. Extracted DNA was stored at ≤−20˚C or used immediately for PCR.

HPV genotyping was performed using the Allplex^TM HPV28 Detection kit on a CFX96^TM Bio-Rad thermocycler, enabling simultaneous detection of 19 high-risk and 9 low-risk HPV genotypes with an integrated internal control. PCR reactions (20 µL) contained 15 µL Master Mix (5 µL HPV28 A or B MOM, 5 µL EM4, and 5 µL EM4 buffer) and 5 µL DNA. Negative, positive, and extraction controls were included. Tubes were sealed and centrifuged to ensure complete contact of the mixture with the tube bottom. Thermal cycling consisted of initial denaturation at 95˚C for 15 min, followed by 45 cycles of 95˚C (3 s), 60˚C (10 s), and 72˚C (10 s), with fluorescence readings at 60˚C, 72˚C, and 83˚C. Samples were considered positive if HPV targets were detected (Ct ≤ 43), and results were validated using internal and external controls.

Sociodemographic and clinical data were collected using structured questionnaires administered by a trained interviewer. A translator (midwife) assisted in cases of language barriers (French or Wolof). Data were recorded in Google Forms, exported to Excel, and analyzed using RStudio (version 2025).

The HPV prevalence observed in this study was low (3.7%), with only one positive case detected, belonging to the HPV 53/82 complex, classified as high-risk but more commonly associated with low-grade cervical lesions. This prevalence is substantially lower than reported in previous African and international studies: 85.9% in a Senegalese cohort of women specifically diagnosed with cervical intraepithelial neoplasia (CIN), a population inherently at higher risk, in which HPV genotypes 16, 18, 45, and 58 predominated [12]; and 37.2% in Kumasi, Ghana, with HPV 52, 56, 35, 18, and 58 being most frequently detected [13].

The low prevalence observed in the present study may be explained by several factors:

Smallsamplesize(n=27), limiting statistical power.Sociodemographicprofileandsexualbehavior: The majority of participants were married and monogamous, reflecting low exposure to high-risk sexual behaviors. Previous studies have reported an association between the number of lifetime sexual partners and the risk of high-risk HPV infection (HPV-HR) [14][15]. Moreover, Shi et al. showed that, according to marital status, HPV prevalence was lowest among married women (29.4%) and highest among single, divorced, or never-married women (>47%) [16]. These findings suggest that sociodemographic characteristics and sexual behavior patterns may contribute to the low prevalence observed in this cohort.Hospital-basedrecruitment, introducing potential selection bias, as most participants were recruited for routine screening (22/27; 81.5%), with fewer women undergoing follow-up for high-grade squamous intraepithelial lesions (HSIL: 2/27; 7.4%) or low-grade squamous intraepithelial lesions (LSIL: 3/27; 11.1%).It is also important to note that cross-sectional studies reflect a single time point and do not capture viral clearance dynamics. Consequently, a low point prevalence does not necessarily indicate a low lifetime risk or cumulative incidence of HPV infection.

HPV53 and HPV82, both possibly carcinogenic to humans (Group 2B), are frequent causes of precancerous lesions but rarely lead to cervical cancer [17]-[19]. Previous studies, including those by Zappacostaetal., suggest that HPV53 may sustain the malignant phenotype induced by HPV16, potentially promoting the progression of high-grade intraepithelial lesions to invasive cervical cancer [20]. HPV-82 can lead to cancer in cases of high viral load or co-infection with a high-risk HPV type, with a reported prevalence of 7 % among 198 women with normal endocervical cytology, according to FayeB.etal., 2020 [21].

The mean age of participants was 44.8 ± 9.6 years (range: 30 - 64 years), consistent with World Health Organization (WHO), Haute Autorité de Santé (HAS), and national guidelines for cervical cancer screening [22][23]. A large proportion of women were married (74.1%), reflecting prevailing local sociocultural norms. Low educational attainment was identified as a major barrier, influencing:

Awareness and understanding of cervical cancer and the importance of screening;Adherence to screening recommendations;Access to healthcare services.

This observation is consistent with previous studies identifying educational level as a key determinant of cervical cancer screening uptake and adherence to preventive programs.

A concerning finding was that 70% of participants had never undergone an HPV test, indicating:

Persistent opportunistic screening;Absence of a structured national screening programme;Insufficient awareness and health education.

The lack of knowledge regarding individual screening status also reflects gaps in medical record traceability, poor communication between healthcare providers and patients, and limited retention of health information. These findings underscore the need for digitalization of medical records to improve follow-up, longitudinal monitoring, and continuity of care.

Traditional risk factors were infrequently reported:

Multiple sexual partners: 33.3% (likely underreported);Unknown HIV status: 40.7%;History of sexually transmitted infections (STIs): 11.1%.

No participant reported HPV vaccination, despite its introduction in the national Expanded Programme on Immunization (EPI) in 2018. The lack of coverage may be explained by the initial target population of girls born between 2004 and 2009, highlighting a gap in vaccination among adult women.

The main limitations of this study include:

Small sample size, limiting generalizability;Cross-sectional design, preventing assessment of viral persistence or temporal dynamics;Enrolment challenges related to participants’ low educational levels;Limited knowledge of medical history among participants, potentially affecting data reliability.

Despite these limitations, the study provides preliminary insights into HPV genotype distribution in a Senegalese hospital-based population and highlights critical areas for improving cervical cancer prevention, including structured screening, health education, and expanded vaccination coverage.