the circulation of respiratory viruses in this region was conducted in 2011, in a different epidemiological context. Since then, no updated data have been published on the dynamics of respiratory infections in Haut-Ogooué. The lack of recent data precludes an up-to-date assessment of the situation, particularly given the potential for significant changes in respiratory virus transmission patterns due to the ongoing impact
A total of 640 individuals were randomly selected from patients who were initially suspected of SARS-CoV-2 infection but tested negative between December 2020 and September 2021. Demographic information (age, sex, and place of residence) and clinical symptoms were obtained from the database of the unit “Emergence Viral Diseases” of the Interdisciplinary for Medical Research of Franceville (CIRMF)”. Nasopharyngeal and oropharyngeal swabs were collected from each participant by trained personnel and placed in viral transport medium. All samples were labeled, registered, and assigned a unique identification number. They were then transported under appropriate conditions to CIRMF, where they were stored at –80°C until further processing.
The programmed thermal cycling involved a temperature of 50°C for 30 minutes, followed by reverse transcription, a temperature of 95°C for 15 minutes, denaturation, a temperature of 45 cycles at 95°C for 15 seconds, 60°C for 34 seconds, and a temperature of 40°C for 10 seconds, respectively. A volume of 20 µl of the mixture (comprising 12.5 µl of 2X one step RT-PCR Buffer, 0.5 µl of 0.1 mm mgso4, 1 µl of 0.04 µg/µl BSA, 1 µl of each sense and antisense primer for each virus, 0.5 µl of each probe and 1 µl of superscript III per sample) was prepared. A volume equivalent to 20 µg/µl bovine serum albumin (BSA), 1 µl of 0.4 µm sense and 1 µl of antisense primers for each virus, 0.5 µl of each probe and 1 µl of superscript III (per sample) was prepared. Finally, 5 µl of the eluate was added to each well of the plate containing the mixture.
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