Genomic Sequencing of SARS-COV-2 in Rwanda: evolution and regional dynamics

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), responsible for coronavirus disease 19 (COVID-19), is a single-stranded positive-sense ribonucleic acid (RNA) virus that typically undergoes one to two single nucleotide mutations per month. COVID-19 continues to spread globally, with case fatality and test positivity rates often linked to locally circulating strains of SARS-CoV-2. Furthermore, mutations in this virus, in particular those occurring in the spike protein (involved in the virus binding to the host epithelial cells) have potential implications in current vaccination efforts. In Rwanda, more than twenty thousand cases have been confirmed as of March 14th 2021, with a case fatality rate of 1.4% and test positivity rate of 2.3% while the recovery rate is at 91.9%. Rwanda started its genomic surveillance efforts, taking advantage of pre-existing research projects and partnerships, to ensure early detection of SARS-CoV-2 variants and to potentially contain the spread of variants of concern (VOC). As a result of this initiative, we here present 203 SARS-CoV-2 whole genome sequences analyzed from strains circulating in the country from May 2020 to February 2021. In particular, we report a shift in variant distribution towards the newly emerging sub-lineage A.23.1 that is currently dominating. Furthermore, we report the detection of the first Rwandan cases of the VOCs, B.1.1.7 and B.1.351, among incoming travelers tested at Kigali International Airport. We also discuss the potential impact of COVID-19 control measures established in the country to control the spread of the virus. To assess the importance of viral introductions from neighboring countries and local transmission, we exploit available individual travel history metadata to inform spatio-temporal phylogeographic inference, enabling us to take into account infections from unsampled locations during the time frame of interest. We uncover an important role of neighboring countries in seeding introductions into Rwanda, including those from which no genomic sequences are currently available or that no longer report positive cases. Our results point to the importance of systematically screening all incoming travelers, regardless of the origin of their travels, as well as regional collaborations for durable response to COVID-19.

Low prevalence of hepatitis C virus RNA in blood donors with anti-hepatitis C virus reactivity in Rwanda.

BACKGROUND: Hepatitis C virus (HCV) is the leading cause of severe liver disease worldwide and is highly endemic in Africa, where it often has nosocomial spread. Little is known on the HCV prevalence, risk for transfusion-transmitted HCV, and circulating genotypes in Rwanda. This study was performed to investigate the prevalence of anti-HCV among blood donors from all regions of the country and genetically characterize identified HCV strains. STUDY DESIGN AND METHODS: Data on anti-HCV reactivity for all 45,061 Rwandan blood donations during 2014 were compiled. Samples from 720 blood donors were reanalyzed for anti-HCV in Sweden. Line immunoassay INNO-LIA HCV and detection of HCV RNA by polymerase chain reaction were used to confirm anti-HCV reactivity. The NS5B and core regions were sequenced and phylogenetic analysis was performed. RESULTS: The anti-HCV prevalence among all first-time blood donors was 1.6%, with the highest occurrence in donors from the eastern region. On further analysis, only 25 of 120 primarily anti-HCV-reactive samples could be confirmed reactive and 15 samples had indeterminate results by INNO-LIA. Confirmed reactivity was more common among females than males (p = 0.03) with no regional difference. Phylogenetic analysis of the sequences showed a predominance of subtypes 4k, 4q, and 4r, with no geographical difference in their distribution. CONCLUSION: The prevalence of anti-HCV among Rwandan blood donors has probably been overestimated previously due to the high rate of nonconfirmable anti-HCV reactivity. Further study of the involved mechanism is needed to avoid loss of blood products and distress for blood donors and other test recipients.

Hepatitis B virus strains from Rwandan blood donors are genetically similar and form one clade within subgenotype A1.

BACKGROUND: Rwanda is a central African country with about 12 million inhabitants. The 1994 genocide against the Tutsi destroyed much of the infrastructure, including the health system. Although this has improved significantly, many challenges remain to be addressed. In this study, the prevalence of serological markers of past and ongoing hepatitis B virus (HBV) infection and HBV vaccine related immunity was investigated in samples from blood donors from all regions of Rwanda. METHODS: The results from hepatitis B surface antigen (HBsAg) analyses of all (45,061) blood donations collected countrywide in 2014 from 13,637 first time and 31,424 repeat blood donors were compiled. Samples from 581 HBsAg negative blood donors were selected for further analysis for antibodies against HBV, anti-HBs and anti-HBc. Additional 139 samples from HBsAg positive donors were analyzed for HBeAg/anti-HBe (132 samples) and for HBV DNA. The S-gene was amplified by PCR, products sequenced, and phylogenetic analysis was performed. RESULTS: HBsAg was found in 4.1% of first time donors with somewhat higher prevalence among those from the Central and Eastern regions than from other parts of the country. Indications of past infection was found in 21% of the HBsAg negative donors, 4.3% had only anti-HBs suggesting HBV vaccination. HBeAg was detected in 28 (21%), anti-HBe in 97 (73%), and both HBeAg and anti-HBe in 4 of 132 HBsAg positive donors. HBV DNA was found in 85 samples, and the complete S-gene was sequenced in 58 of those. Phylogenetic analysis of the sequences revealed that all HBV strains belonged to subgenotype A1, and formed one clade in the phylogenetic tree. In addition, 12 strains from first time donors had a unique 18 amino acid deletion in the N-terminal part of the pre-S2 region. CONCLUSION: This study indicated that the prevalence of hepatitis B is intermediate in Rwanda and that the vaccination coverage is relatively low in young adults. All surveyed Rwandan blood donors were infected with similar subgenotype A1 strains, and a high frequency of those with anti-HBe had detectable HBV DNA. Several strains had in addition a unique pre-S2 deletion, the virulence of which needs to be further studied.