Effect of heat inactivation for the detection of severe acute respiratory syndrome-corona virus-2 (SARS-CoV-2) with reverse transcription real time polymerase chain reaction (rRT-PCR): evidence from Ethiopian study.

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.

Time to recovery and its predictors among adults hospitalized with COVID-19: A prospective cohort study in Ethiopia.

BACKGROUND: Various factors may determine the duration of viral shedding (the time from infection to viral RNA-negative conversion or recovery) in COVID-19 patients. Understanding the average duration of recovery and its predictors is crucial in formulating preventive measures and optimizing treatment options. Therefore, evidence showing the duration of recovery from COVID-19 in different contexts and settings is necessary for tailoring appropriate treatment and prevention measures. This study aimed to investigate the average duration and the predictors of recovery from Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection among COVID-19 patients. METHOD: A hospital-based prospective cohort study was conducted at Eka Kotebe General Hospital, COVID-19 Isolation and Treatment Center from March 18 to June 27, 2020. The Center was the first hospital designated to manage COVID-19 cases in Ethiopia. The study participants were all COVID-19 adult patients who were admitted to the center during the study period. Follow up was done for the participants from the first date of diagnosis to the date of recovery (negative Real-time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCT) test of throat swab). RESULT: A total of 306 COVID-19 cases were followed up to observe the duration of viral clearance by rRT-PCR. Participants' mean age was 34 years (18-84 years) and 69% were male. The median duration of viral clearance from each participant's body was 19 days, but the range was wide: 2 to 71 days. Cough followed by headache was the leading sign of illness among the 67 symptomatic COVID-19 patients; and nearly half of those with comorbidities were known cancer and HIV/AIDS patients on clinical follow up. The median duration of recovery from COVID-19 was different for those with and without previous medical conditions or comorbidities. The rate of recovery from SARS-CoV-2 infection was 36% higher in males than in females (p = 0.043, CI: 1.01, 1.85). The rate of recovery was 93% higher in those with at least one comorbidity than in those without any comorbidity. The risk of delayed recovery was not influenced by blood type, BMI and presence of signs or symptoms. The findings showed that study participants without comorbidities recovered more quickly than those with at least one comorbidity. Therefore, isolation and treatment centers should be prepared to manage the delayed stay of patients having comorbidity.

Sero-prevalence of transfusion transmittable infections: HIV, Hepatitis B, C and Treponema pallidum and associated factors among blood donors in Ethiopia: A retrospective study.

BACKGROUND: Blood transfusion is a therapeutic procedure that has proven to be effective in saving millions of lives. However, its safety is still a crucial issue that needs due attention. Unsafe blood transfusion is one of the sources of transmission for infectious agents. Therefore, the objective of this study was to assess the sero-prevalence of Transfusion Transmittable Infections (TTIs) such as Human immunodeficiency virus (HIV), Hepatitis B Virus (HBV), Hepatitis C virus (HCV), Treponema pallidum, and associated factors among blood donors in Ethiopia. METHODS: A retrospective cross-sectional study design was used to measure the prevalence of transfusion transmittable infections using data collected from 2014 to 2019 in fourteen blood bank facilities in Ethiopia. Screening of HIV, HBV and HCV was done by using the Enzyme-Linked Immunosorbent Assay (ELISA). Presence of Treponema pallidum infection was assessed using rapid plasma reagin (RPR). Records of blood donors were collected using a checklist from Central Blood Bank Laboratory (BBL) electronic database and reviewed. Data was entered, cleaned and analyzed using SPSS version 23. Logistic regression was fitted to identify factors associated with cumulative TTIs positivity, and for each of the transfusion transmittable infection. P value < 0.05 was considered statistically significant. RESULT: A total of 554,954 blood donors in the fourteen blood bank facilities from 2014-2019 was included in the study. The overall sero-prevalence of HBV, Treponema pallidum, HIV and HCV, was 2.4%, 0.9%, 0.4% and 0.4% respectively. The prevalence of TTIs was comparatively higher in 2014, 5.70% and lowest in 2019, 3.40%. The odds of screening HBV in blood donors in age group of 35-39 and 40-44 were 1.2 [1.1, 1.3] and 1.3 [1.1, 1.5] respectively. The odds of screening HCV in blood donors in the age group of 25-34, 35-44 and 45-54 were 1.3 [1.1, 1.5], 1.3 [1.1, 1.7] and 1.7 [1.2, 2.2] respectively. The likelihood of having at least one infection among blood donors was 1.2 [1.1, 1.3] times in male blood donors compared to female. The odds of getting at least one TTI and Treponema pallidum in unemployed blood donors were 2.4 [2.0, 2.8] and 8.1 [6.1, 10.7] respectively. The probability of getting those who have at least one TTIs, HBV and Treponema pallidum were higher in blood donors those who live in Semi Urban and Rural parts of the country than those who live in Urban areas. The odds of having at least one TTI, HBV and HCV in blood donors with mobile mode of blood donation were 1.4 [1.3, 1.6], 1.6 [1.4, 1.8], and 1.6 [1.1, 2.2]. CONCLUSION: The current magnitudes of TTIs are lower when compared to other previous studies conducted in Ethiopia. Older age, male, occupation, donations from mobile sites, residents of semi urban and rural settings were found to be strongly associated with sero-positivity of TTIs. Hence, strict donor screening and testing particularly taking the above factors into consideration is strongly recommended.

Diagnostic accuracy of HIV test kits, Genscreen Ultra and Bioelisa.

PURPOSE: Genetic diversities in different countries affect the performance of HIV test kits. Therefore, WHO recommends evaluation of every HIV test kit in countries' context before its use. Therefore, this study aimed to evaluate the performance of Genscreen ULTRA HIV Ag-Ab and Bioelisa. MATERIALS AND METHODS: The study had used 400 characterized plasma samples obtained from CDC Atlanta bio-bank derived from Africa, USA, and Thailand. RESULTS: Diagnostic performance of both test kits under evaluation was assessed at 95% CI. Genscreen ULTRA HIV Ag-Ab had sensitivity and negative predictive value of 99.5% [95% CI, 97.2-99.9] and the specificity and positive predictive value of 98.5% [95% CI, 95.7-99.7]. Bioelisa HIV test kit had exhibited sensitivity and negative predictive value of 99% [95% CI, 96.4-99.7] and specificity and positive predictive value of 98.5% [95% CI, 95.7-99.7]. Both test kits were able to detect almost all samples with HIV-2, dual infections, and seroconversion. CONCLUSION: Both the test kits were highly sensitive and specific in detecting HIV. However, there are still few samples containing HIV antibody which were not identified by both kits. Therefore, additional screening measures should be done in using these assays for blood transfusion and organ transplantation. In addition, the study can be used as a reference by other African countries.

Evaluation of diagnostic performance of non-invasive HIV self-testing kit using oral fluid in Addis Ababa, Ethiopia: A facility-based cross-sectional study.

BACKGROUND: Human immunodeficiency virus (HIV) testing is critical for early linkage to treatment and care services. However, there is a substantial gap in HIV testing, particularly in resource limited settings due to low accessibility of HIV testing sites, inconvenient testing hours, and concerns about loss of confidentiality. Thus, adopting new strategies such as HIV self-testing (HIVST) could overcome these barriers and increases HIV testing uptake. OBJECTIVE: The aim of this study was to evaluate the diagnostic performance of non-invasive HIVST kit using oral fluid for HIV diagnosis. This study also aimed to assess the ability of clients in interpretation of OraQuick HIVST results. METHOD: Between December 2017 and February 2018, a total of 400 study participants were enrolled into the study to assess a diagnostic accuracy of Oral fluid-based HIVST kit (OraQuick) in 15 public health facilities in Addis Ababa, Ethiopia. Participants were provided with instructions and visual aids on how to perform HIVST and interpret results. They also underwent a blood-based rapid HIV antibody test as per the current national algorithm. The results of HIVST were interpreted independently by the participants and respective health care workers (HCWs). The sensitivity, specificity, positive predictive value (PPV), Negative predictive value (NPV) and inter-rater agreement of the test were computed. RESULTS: Out of 200 participants who tested positive on the national algorithm testing, oral fluid-based self-testing was positive in 199 (99.5%), false negative in 1 (0.5%). Of 200 participants who tested negative on the national algorithm testing, self-testing was negative in 200 (100%). There were no false positive and invalid tests. The sensitivity and specificity of the OraQuick HIVST were 99.5% (95%CI: 97.26-99.99) and 100% (95%CI: 98.18-100.0), respectively. The overall agreement between the two tests was high (κ value = 0.995). The PPV and NPV of OraQuick test were 100% and 99.5% (95%CI: 96.59-99.93) respectively. CONCLUSION: This study showed a high diagnostic performance of OraQuick HIV self-test and suggests that OraQuick HIVST kit has a potential to be used for HIV testing in Ethiopia along with the national algorithm.