Study design, area, and period

This hospital-based cross-sectional study was conducted at Addis Zemen Primary Hospital (AZPH) from April to July 2023. AZPH is found in Northwestern Ethiopia, located in the South Gondar zone of the Amhara national regional state, on the road connecting Bahir Dar and Gondar. Addis Zemen is a town located at a latitude and longitude of 12o7’N37o47’E and at an altitude of 1975 m above sea level. It is the administrative center of the Libo Kemkem district and is located 102 km from Bahir Dar and 645 km from Addis Ababa.

Sample size determination and sampling technique

The sample size for this study was determined by the formula for single population proportion formula with 50% prevalence, 95% confidence level, and 5% margin of error, and the sample was collected using a convenient sampling technique until a total of 384 samples were enrolled.

Eligibility criteriaInclusion criteria

All patients with gastrointestinal complain (Abdominal pain, diarrhea, constipation, nausea, vomiting, bloating etc.) who are voluntary to participate in the study were included.

Exclusion criteria

Patients who have taken anti helminthic and anti-protozoal drugs within the past two weeks before data collection were excluded.

Dependent and independent variables

Dependent variables in this study were intestinal parasite and H. pylori coinfection status. Independent variables included age, sex, educational status, occupation, fingernail status, residence, family size, family history of H. pylori, and source of drinking water.

Operational definitions

Family size was determined as;

*Small: Family consisting of one to four members.

*Large: Family consisting of five or more members.

Data collection and processing Sociodemographic data collection

The sociodemographic data of patients suspected of intestinal parasitosis and/or gastritis that came to the laboratory upon stool examination request were collected using a structured questionnaire via face-to-face interviews.

Stool sample collection and laboratory investigation

The patients were informed to pass stool samples directly inside a plastic cup with a compacted lid. All specimen containers were labeled appropriately with the patient’s name, medical registration number, age, sex, and date of sample collection. About 10–20 g of stools or 5–6 spoons full of watery stools were collected. After the sample was collected in a clean, dry, leak-proof container, intestinal parasites and H. pylori were examined using saline wet mount and serological H. pylori stool antigen tests (manufactured by Guangzhou Wondfo Biotech Co., Ltd.), respectively.

Direct wet mount

A matchstick head-sized stool sample was placed over a glass slide and mixed with one drop of normal saline solution. The slides were shielded with a glass cover slip and examined for the presence of parasite eggs, larvae, trophozoites, and cysts at 10× and 40 × magnifications with Olympus Cx21FS1 Microscope (OLYMPUS CORPORATION, TOKYO JAPAN, made in Philippines).

H. pylori stool antigen test

The stool collection apparatus was opened and a collection stick was used to prick the stool sample. Then, the collection stick containing the stool sample was placed back into the sample collection tube and shaken vigorously to mix with the sample solution. The test kit (Wondfo one-step H. pylori feces kit, catalog number Wo28, manufactured by Guangzhou Wondfo Biotech Co., Ltd.,) was removed from the foil pouch by tearing at the notch and placed on a level surface. When the sample collector was held upright, the tip of the collector broke at the breakpoint. Three drops of the sample solution were dispensed inside the sample well and allowed to stand for 15 min. The results were considered positive when rose-pink bands were visible in both the control and test regions. The results were reported as negative when a rose-pink band was visible only in the control region.

Data quality assurance

Prior to the start of the data collection, training was provided to the data collectors. Patients were informed accordingly before stool sample collection. Inappropriate samples, such as those with insufficient amounts, were rejected and recollected to reduce errors in results. The reliability of H. pylori stool antigen test was evaluated by implementing daily quality control measures, using known positive and negative samples. About 5% (20) of the samples were randomly checked by experienced microscopists to ensure the consistency of results for intestinal parasites.

Data analysis

The questionnaire data on sociodemographic characteristics and associated factors were checked for completeness and analyzed using the Statistical Package for Social Sciences version 20 (SPSS 20). Descriptive statistics (frequencies and percentages) were used to display the results of the variables. Bivariate logistic regression was used to select variables, and multiple logistic regression was used to determine the association between independent and outcome variables. Variables with p-value < 0.25 in the bivariate logistic regression were selected for multivariate logistic regression analysis. Adjusted Odds ratios (AORs) with 95% confidence intervals were calculated and p-values < 0.05 were considered statistically significant.

Ethical considerations

This study was reviewed by Department of Medical Laboratory Science, Debre Tabor University research review committee with the approval number (157/2023). Written informed consent was obtained from the adult study participants. Written informed assent was obtained from the parents and guardians of the study participants aged below 15 years old. The results were reported to the concerned physicians and patients with positive results were treated accordingly.