Study setting and population
This study was conducted in public health facilities of Shashemene town, Oromia Regional State, Ethiopia, 250 Km from Addis Ababa, from February 1 to March 30, 2022. The town has an estimated total population of 382,216 in 2022(Shashemene town population statistics). Based on town health office report, the town has 2 governmental hospitals, 1 private hospital, 4 functional public health centers and 1 private health center, of which currently 3 public health facilities are providing cervical cancer screening service such as HPV screening.
All WLWH who come to Shashemene town public health facilities ART clinic were source population. All WLWH who come to Shashemene town public health facilities ART clinic and tested for HPV during study period were our study population, whereas women who had hysterectomy done and impossibility of obtaining samples were excluded from the study
Sample size determination
Sample size was determined using single population proportion formula with the assumption of 50% proportion of HR HPV among WLWH was used (since there was no previous study in our setting) and 95% confidence level (CL) and 5% of marginal error(d). Finally, by adding 10% of non-response rate the total sample size was estimated to be 422.
Sampling procedure
All public Health facilities currently providing HPV testing in the town namely Shashemene comprehensive specialized Hospital (SCSH), Melka Oda General Hospital (MOGH), and Abosto health center (Hc) were selected. Based on the number of eligible women and women appointed during the study period, the total sample size was allocated proportionally to each health facility. A study subject was selected using systematic random sampling technique. K value was calculated for each health facility and average k value (k = 2) was taken
Data collection Tools and procedure
A questionnaire was adapted from different literature's of similar studies [15–21] and adapted to the current aim of the study. The prepared questionnaire included information on socio-demographic characteristics, sexual and reproductive health factors, behavioral factors, and clinical related factors. The questionnaire was first prepared in English and the translated to Afan Oromo and Amharic language by two independent translators who are health professionals and can speak both Afan Oromo and Amharic language(local language of the study area) by two independent translators who are health professionals and can speak both languages. It was back translated to English by independent translator to make sure the questions are clear and to check for consistency. The data was collected by 6 trained health professionals currently working on CC screening using face-to-face interviews. Patient’s chart was reviewed to retrieve medical information like; CD4 count, viral load, length of being on ART and WHO clinical staging of the patients from patient card after the result is returned.
Laboratory procedure
A sample of cells for Hr HPV test was collected by a trained health care provider after the patient was placed in a dorso lithotomy position by inserting a swab deep into the cervix and rotating as directed. The health care provider then places the swab in the sleeve. The closed sleeve was kept at room temperature and sent to Shashemene regional laboratory which is found inside Shashemene comprehensive specialized Hospital at ambient temperature in weekly or bi-weekly shipment. At the testing laboratory the sample was kept at 4°C until extraction (within 3 weeks). DNA extracts are stored temporarily at 4 ºC or for long-term storage, in -80ºC freezer. Then, DNA extraction and high-risk HPV detection from collected cervical and vaginal swabs was conducted. DNA extraction was performed with a modified protocol and the commercial QIA amp kit (Qiagen, Valencia, CA) and the presence of high-risk HPV in the extracted patient DNA was determined with the Cobas 4800 HPV Test. The Cobas 4800 HPV test identifies the presence of HPV type 16, 18 and other Hr HPV (12 genotypes of Hr HPV).
Data quality management
The quality of data was assured by applying a properly designed and pre-tested questionnaire. The tool was pre-tested on five percent (5%) of the sample size at Dodola General Hospital before the actual data collection to establish its ability to elicit relevant information. Regular supervision and follow-up were made by supervisor. In addition, regular check-up for completeness and consistency of the data was made on daily basis and checking of questionnaire consistency was made. Incomplete questionnaires, was corrected on spot.
five percent (5%) of known sample was taken from Hawasa regional laboratory and rechecked at Shashemene regional laboratory. regular check-up for proper labeling of collected sample was made. Again in testing unit, expiry date and validity of test kit was regularly checked. For sample tests invalid by the Cobas 4800 HPV test, the DNA from that sample was retested one time to obtain valid results. For repeated invalid result, the final result was recorded as invalid. Improperly labeled sample and invalid result was discarded and considered as non- response rate.
Variable measurement
Our dependent variable is presence or absence of high risk human papilloma virus. It is a dichotomous variable and labeled as “yes” for High risk HPV and “no” for others (Low risk HPV and not found) as defined under operational definition.
Our independent variables include age (15–24, 25–34, 35–44 and ≥ 44 years), ethnicity (Oromo, Amhara, others (Wolayita, Tigre)), religion (Muslim, Orthodox, Protestant, others(Catholic)), Educational status(illiterate, only reading and writing, primary school, secondary school, and college and above)), marital status (married, widowed and others (single, divorced)), residence (urban, and rural), occupation (government employee, house wife, merchants and others (farmer, daily labourer)), husband’s educational level (no formal education, primary school, secondary school, and college and above)), husband’s occupation (farmer, merchant, drive, daily labourer and government employee), ever heard about cervical cancer screening, source of information (health professional, TV/Radio and neighbors), previous history of CC screening, type of screening (VIA and others (pap smear, HPV screening)), history of chronic disease (hypertension, diabetes), history of STI, years of ART Attendance (< 2 years and ≥ 2 years), previous change of ART regimen, baseline and end-line CD4 count (< 200cell/mm3 and > 200cell/mm3), baseline and end-line HIV viral-load (< 50 copies/ml and ≥ 50 copies/ml), WHO clinical stage (stage 1 and others (stage 2, 3, and 4)), cigarette smoking, drinking alcoholic drinks (beer, wine and alcohols), chewing chat, taking any other drug (other than cigarettes, alcohol or khat), number of birth (≤ 1, and > 1), age at first sexual intercourse (< 18 years and ≥ 18 years), number of life time sexual partner (1 and > 1), number of her husband’s life time sexual partner (1 and > 1), use of modern contraceptive methods and history of abortion
Data processing and Analysis
The collected data was checked; coded and entered into Epi info version 7.2.5 and exported to SPSS version 24.0 for further analysis. Descriptive statistics like frequency and cross-tabulation were carried out to summarize data. Bivariate logistic regression analysis were carried out to check the association of individual variables with the outcome variable. Those variables with significance level p-value < 0.2 in bivariate analysis was entered into multivariable logistic regression model for further analysis and the backward stepwise (likelihood ratio) were carried out to adjust for the confounding variables. The Hosmer–Lemeshow goodness-of-fit statistic was used to assess whether the model is fit ( p-value 0.396). Odds ratios (OR) and the corresponding 95% confidence intervals (95% CI) were calculated to assess the degree of association between independent and dependent variables. Finally, the variables with a p-value less than 0.05 were considered as statistically significant.
Operational definition
High risk human papilloma virus (HR HPV)
is a type of HPV (HPV type 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68) that can cause cervical cancer and other types of cancer, such as cancers of the anus, vagina, vulva, penis, and oropharynx. Chronic infection with Hr HPV can lead to cell changes that, if not treated, may become cancer [27].
Low risk human papilloma virus (LR HPV)
- is a type of HPV (HPV type 6, 11, 42, 43, 44, 54, 61, 70, 72, and 81) that can cause skin warts and respiratory papillomatosis, it does not cause and many of them go away on their own without treatment [27].