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Background: Even though highly active antiretroviral therapy (HAART) for HIV (Human Immune Deficiency) patients has considerably improved viral load suppression, more people still struggle to reduce viral loads. The aim of the study was to determine the associated factors of virological failure in HIV patients on antiretroviral therapy (ART) in the Upper East Region of Ghana. Methods: A retrospective cross-sectional study involving 366 participants aged 15 years and above who were on HAART for six (6) months or longer with viral load results in 2020. Bivariate and multiple logistic regression analyses were conducted to identify the determinants of virological failure among HIV patients at 95% confidence interval (C.I.) with a significant level pegged at a p value less than 0.05. Results: The prevalence of viral load failure was 47.0% and adherence to antiretroviral therapy was 62.6% among patients on HAART. The significant predicators of virological failure were basic education (AOR (adjusted odds ratio) = 7.36, 95% C.I = 4.91–59.71), High school/Vocational /Technical education (AOR = 4.70, 95% C.I. = 1.90–9.69), monthly salary/income ˂ GHS 375.00 (AOR = 7.20, 95% C.I. = 1.73–29.95), duration on ART for ˂1 year (AOR = 0.27, 95% C.I. = 0.10–0.75), ART regimen (Tenofovir + Lamivudine + Efavirenz) (AOR = 3.26, 95% C.I. = 1.95–11.25), 3–5 times missed medication per month (AOR = 2.86, 95% C.I. = 1.34–6.08) and ≥6 missed medication per month (AOR = 23.87, 95% C.I. = 10.57–53.92). Conclusion: Educational status, salary/income, ART duration, ART combination regimen, and number of doses missed were statistically significantly associated with virological failure in patients on antiretroviral therapy. The majority of the respondents adhered to ART, which led to moderate viral load suppression but lower than the target for 2020. There is the need to strengthen the ongoing accelerated social behavior change communication among patients on ART to enhance adherence in order to attain the new UNAIDS target of 95% viral load suppression by 2030 in the Upper East Region of Ghana.

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