Habarugira, G. | Search bioRxiv

bioRxiv2024

Application of chimeric antigens to paper-based diagnostics for detection of West Nile virus infections of Crocodylus porosus: a novel animal test case

Laboratory-based diagnostics like plaque reduction neutralization tests (PRNT) and ELISA are commonly used to detect seroconversion to flavivirus infections. However, faster, qualitative screening methods are needed for quicker diagnosis and better patient outcomes. Lateral flow assays (LFAs) can provide rapid results (5-15 mins) at the point-of-care, yet few commercial flavivirus antibody detection LFAs are available. We developed an LFA using novel chimeric viral antigens produced by genetically modifying the mosquito restricted Binjari virus (BinJV) to display the outer virion proteins of pathogenic viruses such as West Nile virus (WNV). The BinJV chimeric platform offers various advantages for diagnostic assay development, including rapid construction of new chimeras in response to emerging viral variants, safe, scalable antigen manufacturing, and structural indistinguishability to the wild-type pathogenic virion. As a demonstration of feasibility, we applied chimeric WNV (BinJV/WNV) antigen to LFA as the capture/test line reagent for detection of seroconversion of crocodilians to WNV - a virus affecting crocodilians on multiple continents. We verified the antigenic conservation of the chimera when applied to the LFA detection surface using monoclonal antibodies. Using well-characterised sera (n=60) from WNV seropositive or flavivirus naive Australian saltwater crocodiles (Crocodylus porosus), we illustrated 100% sensitivity and specificity, with results achieved in less than 15 minutes. The LFA further accurately detected seroconversion in animals experimentally infected with WNV. This qualitative screening method can be performed both inside and outside of a laboratory, and the assay design will guide the optimization of similar tests for vector borne virus infection detection in both humans and other animals.

biochemistry↗

bioRxiv2020

Magnaporthe oryzae populations in Sub-Saharan Africa are diverse and show signs of local adaptation

Rice blast caused by Magnaporthe oryzae is one of the most economically damaging diseases of rice worldwide. The disease originated in Asia but was detected for the first time in Sub-Saharan Africa (SSA) around 100 years ago. Despite its importance, the evolutionary processes involved in shaping the population structure of M. oryzae in SSA remain unclear. In this study, we investigate the population history of M. oryzae using a combined dataset of 180 genomes. Our results show that SSA populations are more diverse than earlier perceived, and harbor all genetic groups previously reported in Asia. While M. oryzae populations in SSA and Asia draw from the same genetic pools, both are experiencing different evolutionary trajectories resulting from unknown selection pressures or demographic processes. The distribution of rare alleles, measured as Tajimas D values, show significant differences at the substructure level. Genome-wide analysis indicates potential events of population contraction strongly affecting M. oryzae in SSA. In addition, the distribution and haplotype diversity of effectors might suggest a process of local adaptation to SSA conditions. These findings provide additional clues about the evolutionary history of M. oryzae outside the center of origin and help to build customized disease management strategies.

genomics↗