Background: Coronavirus disease 2019 (COVID-19) has been a major public health importance and its specimen needs to be handled safely due to concerns of potential transmissibility to health care workers. Heat inactivation of the sample before nucleic acid isolation might permit safe testing processes. Hence, it is important to assess the effect of heat inactivation on SARS-CoV-2 RT-PCR detection in resource limited settings. Methods: An experimental study was conducted at Ethiopian Public Health Institute (EPHI) from September 25 to October 15, 2020. A total of 188 Oro-pharyngeal swabs were collected from COVID-19 suspected cases, referred to EPHI for SARS COV-2 testing. One batch of the sample was inactivated at 56 °C heat for 30 min, and the other batch was stored at 4 °C for a similar period of time. RNA extraction and detection were done by DAAN Gene kit protocols. Abbott m2000 RT-PCR was used for amplification and detection. Data analysis was done by using SPSS version 23.0; Chi-square and Pearson correlation test for qualitative and semi-quantitative analysis were used. p-value < 0.05 was considered as statistically significant. Results: Out of 188 total samples, 119 (63.3%) were positive and 69 (36.7%) were negative in the non-inactivated group. While, 115 (61.2%) of samples were positive and 73 (38.8) were negative in heat inactivated sample batch. Rate of positivity between groups did not have statistically significant difference (p > 0.05). The mean Cycle threshold (Ct) value difference between the two groups of ORF1a/b gene and N gene was 0.042 (95% CI − 0.247–0.331; t = 0.28; p = 0.774) and 0.38 (95% CI 0.097–0.682; t = 2.638; p = 0.010) respectively. Conclusion: Heat inactivation at 56 °C for 30 min did not affect the qualitative rRT-PCR detection of SARS-CoV-2. However, the finding showed that there was statistically significant Ct value increment after heat inactivation compared to untreated samples. Therefore, false-negative results for high Ct value (Ct > 35) samples were found to be the challenge of this protocol. Hence alternative inactivation methods should be investigated and further studies should be considered.

Since Coronavirus Disease-2019 (COVID-19) outbreak was reported, many commercial Nucleic Acid Amplification Tests (NAAT) have been developed all over the world, and it has been the standard method. Even though several assays were rapidly developed and applied to laboratory diagnostic testing, the performance of these assays was not evaluated in different contexts. Thus, this study aimed to assess the performance of Abbott SARS-CoV-2, Daan Gene, BGI and Sansure Biotech assays by using Composite Reference Standard (CRS). The study was conducted at the Ethiopian Public Health Institute (EPHI) from December 1 to 30/2020. Of the 164 nasopharyngeal samples were extracted by using a QIAamp RNA mini kit and Abbott DNA sample preparation system. Out of 164 samples, 59.1% were positive and 40.9% were negative by CRS. Sansure Biotech positivity was significantly low compared to CRS (p < 0.05). The overall agreement of the four assays compared to CRS was 96.3–100%. The performance of the four assays had almost comparable diagnostic performance, except for a low positive rate of Sansure Biotech assay. Hence, Sansure Biotech assay [Research Use Only (RUO)] needs further verification on its use in Ethiopia. Finally an additional study should be considered for evaluating assays with respective manufacturer claims.

HIV drug resistance (HIVDR) presents a significant challenge to antiretroviral therapy (ART) success, particularly in resource-limited settings like Ethiopia. This cross-sectional study investigated viral suppression rates and resistance patterns among patients on second-line ART across 28 Ethiopian health facilities. Blood samples collected from 586 participants were analyzed to measure CD4 count and viral load and assess HIVDR in patients experiencing virological failure (VF) (viral load ≥ 1000 copies/mL). Demographic and clinical data were analyzed using logistic regression to identify factors associated with VF. Results showed that 13.82% of participants experienced VF, with 67.57% of genotyped samples exhibiting at least one drug resistance mutation. Resistance to nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), and protease inhibitors (PIs) was detected in 48.64%, 64.86%, and 18.92% of cases, respectively. Dual-class resistance was identified in 48.64% of patients, while triple-class resistance was detected in 18.92%. VF was more likely among students and those with CD4 counts below 200 cells/mm³, but less likely in patients on second-line treatment for 12 months or more. Our findings highlight a substantial HIVDR burden among patients on second-line ART with VF, emphasizing the need for comprehensive HIV care, including adherence support, regular viral load monitoring, and HIVDR testing.

Objective: Despite global progress in reducing HIV-related morbidity and mortality, preventable deaths persist, particularly in sub-Saharan Africa. Ethiopia has achieved near-universal antiretroviral therapy (ART) coverage, yet survival outcomes and predictors of mortality in the era of ‘test and treat’ remain underexplored. This study examined survival rates and predictors of mortality among people living with HIV (PLHIV) on ART in Ethiopia between 2016 and 2023. Design: We conducted a retrospective open cohort study. Setting: 65 public health facilities across Ethiopia. Kaplan-Meier survival analysis and multivariable Cox proportional hazards regression were applied. Statistical significance was set at p<0.05. Participants: The analysis included 33 416 PLHIV enrolled in care and treatment from 2016 to 2023. In the current study, baseline is defined as the start date of ART treatment. Primary and secondary outcome measures: The primary outcome was survival status at the end of follow-up, defined as death versus alive. Result: Among 33 416 PLHIV, 2659 deaths (8.6%) were documented during 85 972 person-years of follow-up, yielding a mortality incidence rate of 3.09 per 100 person-years. Most deaths occurred within the first 12 months of ART initiation. Independent predictors of mortality included age (≥35 years), bedridden (AHR 2.46, 95% CI 1.78 to 3.40) and ambulatory status (AHR 2.33, 95% CI 1.84 to 2.95), WHO stage IV (AHR 1.40, 95% CI 1.02 to 1.91), having opportunistic infections (AHR 1.41, 95% CI 1.13 to 1.76) and baseline CD4 <200 cells/mm³ (AHR 3.08, 95% CI 2.31 to 4.09). Receipt of tuberculosis preventive therapy (TPT) was protective (AHR 0.36, 95% CI 0.30 to 0.43). Conclusion: The results show that many people appear to start ART late. Factors such as impaired functional status, the presence of opportunistic infections and older age appear to have further compounded risk, while TPT appears to be strongly protective. Improving early diagnosis, timely ART initiation and TPT uptake could further support better outcomes.

Objectives: This study aimed to determine the prevalence of detectable HIV-1 viral load and associated factors among pregnant and lactating women on Dolutegravir-based and non-Dolutegravir-based regimens. Methods: HIV-positive pregnant and breastfeeding women enrolled in a cross-sectional study design incorporating Dolutegravir versus non-Dolutegravir group for comparison purposes, and the study was conducted from September 2022 to March 2023. In total, 684 women receiving antiretroviral treatment (456 on Dolutegravir-based and 228 on non-Dolutegravir-based regimens) were enrolled across 16 prevention of mother-to-child transmission clinics. An undetectable viral load was defined as <20 RNA copies/mL. Univariable and multivariable regression models were used to determine the associations of the detectable viral load with factors such as sociodemographic, clinical, obstetric, and laboratory parameters. Results: The prevalence of detectable viral load on a non-Dolutegravir-based regimen was 32.9% and on a Dolutegravir-based regimen group was 19.7%. The overall prevalence was 24.1% (95% CI: 20.9–27.6). Compared with non-Dolutegravir-based regimens, Dolutegravir-based regimens were associated with 69% (AOR: 0.31; 95% CI: 0.16–0.61) lower risk of having a detectable viral load. The median viral load was significantly lower in women on a Dolutegravir-based regimen (1.66 log copies/mL) compared to those on a non-Dolutegravir-based regimen (2.11 log copies/mL; p < 0.001). A lower level of adherence was associated with 2.33 times (AOR: 2.33; 95% CI: 1.06–5.12) higher odds of having a detectable viral load and women who had a WHO clinical stage II and above were 2.71 times (AOR: 2.71: 95% CI: 1.30–5.64) more vulnerable to developing a detectable viremia than their counterparts. Conclusion: Despite the scale-up of Dolutegravir-based regimens, the overall prevalence of detectable viremia was 24.1%, with a higher prevalence of 32.9% among women on non-Dolutegravir-based regimens indicating a programmatic challenge that could affect mother-to-child transmission outcomes. Strengthening adherence monitoring and maintaining World Health Organization clinical stage I is crucial for achieving undetectable viral load levels in pregnant and breastfeeding women.

Since coronavirus disease-2019 (COVID-19) outbreak was reported, many commercial Nucleic Acid Amplification Testing (NAAT) assays have been developed all over the world, and it has been the standard method. Though, a number of assays were rapidly developed and applied to clinical testing, performance of these assays were not evaluated in different contexts. Thus, this study was aimed to assess the performance of common assays (Abbott SARS-CoV-2, DaanGene, BGI and Sansure Biotech) available in Ethiopia by using combined reference results (CRR). The study was conducted at Ethiopian Public Health Institute (EPHI) from December 1 to 30/2020. A total of 164 nasopharyngeal samples were extracted by using QIAamp® RNA mini kit and Abbott DNA sample preparation system. Out of 164 samples, 59.1% was positive and 40.9% was negative by CRR. Sansure Biotech positivity was significantly low compared to CRR (p<0.05). The overall agreement of the four assays compared to CRR was 96.3% to 100%. The performance of four assays had almost comparable diagnostic performance, except a low positive rate of Sansure Biotech assay. Hence, Sansure Biotech assay (RUO) needs further verification on its use in Ethiopia. Finally, an additional study should be considered for the evaluation of assays with respective manufacturer claims.