Background: HIV poses significant challenges for vaccine development due to its high genetic mutation and recombination rates. Understanding the distribution of HIV subtypes (clades) across regions and populations is crucial. In this study, a systematic review of the past decade was conducted to characterize HIV-1/HIV-2 subtypes. Methods: A comprehensive search was performed in PubMed, EMBASE, and CABI Global Health, yielding 454 studies from 91 countries. Results: Globally, circulating recombinant forms (CRFs)/unique recombinant forms (URFs) accounted for 29% of HIV-1 strains, followed by subtype C (23%) and subtype A (17%). Among studies reporting subtype breakdowns in key populations, 62% of HIV infections among men who have sex with men (MSM) and 38% among people who inject drugs (PWIDs) were CRF/URFs. Latin America and the Caribbean exhibited a 25% increase in other CRFs (excluding CRF01_AE or CRF02_AG) prevalence between 2010–2015 and 2016–2021. Conclusions: This review underscores the global distribution of HIV subtypes, with an increasing prevalence of CRFs and a lower prevalence of subtype C. Data on HIV-2 were limited. Understanding subtype diversity is crucial for vaccine development, which need to elicit immune responses capable of targeting various subtypes. Further research is needed to enhance our knowledge and address the challenges posed by HIV subtype diversity.

Arboviruses often cause widespread morbidity in children in endemic regions. Data on the burden of arboviruses in Kenyan children are limited. This study was performed to determine the seroprevalence of yellow fever (YFV), dengue (DENV), West Nile (WNV), and chikungunya (CHIKV) viruses among children 1-12 years of age at two health facilities in Teso South Sub-County in Western Kenya. In a hospital-based cross-sectional survey, a questionnaire was used to collect socio-demographic information. Serum drawn from the children was tested for IgA/IgM/IgG serocomplex antibodies to selected arboviruses using indirect ELISA and plaque reduction neutralization tests. A total of 182 (27.7%) of the 656 participants tested were positive for any arbovirus antibody. Of these, 4.4% (29/656) tested positive for YFV, 9.6% (62/649) for WNV, 5.6% (36/649) for CHIKV, 1.4% (5/368) for DENV1, 9% (59/656) for DENV2, and 19.7% (40/203) for DENV3. Neutralizing antibodies to CHIKV were found in 77.8% (42/54) of participants, to YFV in 15.8% (3/19), to DENV2 in 58% (29/50), and to WNV in 8% (1/55). Sex, age, urban residence, schooling, and lack of vaccination were associated with arbovirus exposure. This study confirmed that children under 12 years of age in Teso South Sub-County are exposed to ongoing arbovirus infections early in life.

CD4+ T cell enumeration is used to determine eligibility for antiretroviral therapy (ART) and to monitor the immune status of HIV-positive patients; however, many patients do not have access to this essential diagnostic test. Introducing point of care (POC) testing may improve access. We have evaluated Alere’s PIMA™, one such POC device, against conventional CD4+ testing platforms to determine its performance and validity for use in Kenya. In our hands, Alere PIMA™ had a coefficient of variability of 10.3% and of repeatability of 175.6 cells/µl. It differed from both the BD FACSCalibur™ (r² = 0.762, mean bias −64.8 cells/µl), and the BD FACSCount™ (r² = 0.874, mean bias 7.8 cells/µl). When compared to the FACSCalibur™ at a cutoff of 350 cells/µl, it had a sensitivity of 89.6% and a specificity of 86.7% in those aged 5 years and over (Kw = 0.7566). With the BD FACSCount™, it had a sensitivity of 79.4% and a specificity of 83.4% in those aged 5 years and over (Kw = 0.7790). The device also differed from PARTEC Cyflow™ (r² = 0.781, mean bias −24.2 cells/µl) and GUAVA™ (r² = 0.658, mean bias −0.3 cells/µl) platforms, which are used in some facilities in Kenya. We conclude that with refinement, Alere PIMA™ technology has potential benefits for HIV-positive patients. This study highlights the difficulty in selecting the most appropriate reference technology for technical evaluations.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease that was first reported in China in 2011. It is caused by SFTS virus (SFTSV) which is a member of the Phlebovirus genus in the Bunyaviridae family. SFTSV has been classified as a BSL3 pathogen. There is a need to develop safe and affordable serodiagnostic methods for proper clinical management of infected patients. The full length nucleocapsid (N) gene of SFTSV Yamaguchi strain was amplified by RT-PCR and cloned to an expression vector pQE30. The recombinant (r) SFTSV-N protein was expressed by using Escherichia coli (E. coli) expression system and purified under native conditions. rSFTSV-N protein based indirect IgG and IgM enzyme linked immunosorbent assay (ELISA) systems were established to detect specific human IgG and IgM antibodies, respectively. One hundred fifteen serum samples from clinically suspected-SFTS patients were used to evaluate the newly established systems and the results were compared with the total antibody detecting sandwich ELISA system. The native form of recombinant (r) SFTSV-N protein was expressed and purified. Application of the rSFTSV-N protein based indirect IgG ELISA to the 115 serum samples showed results that perfectly matched those of the total antibody sandwich ELISA with a sensitivity and specificity of 100 %. The rSFTSV-N protein based indirect IgM ELISA missed 8 positive samples that were detected by the total antibody sandwich ELISA. The sensitivity and specificity of rSFTSV-N-IgM capture ELISA were 90.59 and 100 %, respectively. The rSFTSV-N protein is highly immunoreactive and a good target for use as an assay antigen in laboratory diagnosis. Its preparation is simpler in comparison with that used for the total antibody sandwich system. Our rSFTSV-N protein-based IgG and IgM ELISA systems have the advantage of distinguishing two types of antibodies and require small volume of serum sample only. They are safe to use for diagnosis of SFTS virus infection and especially fit in large-scale epidemiological investigations.

Yellow fever (YF) is an acute hemorrhagic viral infection transmitted by mosquitoes in Africa and South America. The major challenge in YF disease detection and confirmation of outbreaks in Africa is the limited availability of reference laboratories and the persistent lack of access to diagnostic tests. We used wild-type YF virus sequences to generate recombinant envelope protein in anEscherichia coliexpression system. Both the recombinant protein and sucrose gradient-purified YF vaccine virus 17D (YF-17D) were used to immunize BALB/c mice to generate monoclonal antibodies (MAbs). Eight MAbs were established and systematically characterized by indirect enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and immunofluorescence assay (IFA). The established MAbs showed strong reactivity with wild-type YF virus and recombinant protein with no detectable cross-reactivity to dengue virus or Japanese encephalitis virus. Epitope mapping showed strong binding of three MAbs to amino acid positions 1 to 51, while two MAbs mapped to amino acid positions 52 to 135 of the envelope protein. The remaining three MAbs did not show reactivity to envelope fragments. The established MAbs exert no neutralization against wild-type YF and 17D viruses (titer of <10 for both strains). The applicability of MAbs 8H3 and 3F4 was further evaluated using IgM capture ELISA. A total of 49 serum samples were analyzed, among which 12 positive patient and vaccinee samples were correctly identified. Using serum samples that were 2-fold serially diluted, the IgM capture ELISA was able to detect all YF-positive samples. Furthermore, MAb-based antigen detection ELISA enabled the detection of virus in culture supernatants containing titers of about 1,000 focus-forming units.

Background: Tsetse flies are the cyclical vectors of both human and animal diseases. Kenya’s commitment to eradicate tsetse and trypanosomiasis dates to the 1980s through various control approaches which were spearheaded by the African Union. The aggressive control programmes together with climatic, land-use, and socio-economic changes immensely contributed to the reduction of African trypanosomiasis. Since 2012, Kenya has not recorded a case of human trypanosomiasis. However, African animal trypanosomiasis remains a major challenge to livestock production in 38 out of 47 counties. We aimed to determine the prevalence of tsetse flies and trypanosome infection rate and to build the capacity of small-holder livestock producers in vector control activities in Busia county.Methods: This cross-sectional study was conducted between May 2018 and December 2018 in Busia county, a beneficiary of the previous African Union-led trypanosomiasis and tsetse control initiatives. Odour-baited biconical traps were deployed for 48 h in five sampling areas. Captured tsetse flies were analysed by microscopy for trypanosome infections. Additionally, training and field demonstrations were conducted as part of capacity building to enhance participation of small-holder livestock producers in tsetse control activities.Results: A total of 94 tsetse flies mainlyGlossina fuscipes fuscipeswere captured from the five sampling areas. The apparent fly densities range from 0.08 to 1.55 tsetse per trap per day. Additionally, 75 biting flies mainlyStomoxysspp. were also trapped. An overall tsetse infection rate of 1.39% and 4.17% was observed forTrypanosoma congolenseandTrypanosoma vivax, respectively. Regarding capacity building, a total of 26 small-holder livestock producers were trained on tsetse and trypanosomiasis control activities. Out of which, five were selected as focal persons and were further trained on integrated vector management techniques and tsetse survey methods.Conclusions: Our findings revealed the existence of trypanosome-infected tsetse flies which could potentially spread to other parts of the county. Training of small-holder livestock producers in tsetse and trypanosomiasis control activities should be supported and integrated in the county animal health and veterinary services. Given the observed low tsetse densities and trypanosome infection rates, the elimination of trypanosomiasis in Busia county is feasible.