Fig 1.
Map of Sri Lanka showing the areas of origin of human samples used for the study.
The region where rodent-borne virus sequences used in this study were previously identified is also indicated. The basemap of Sri Lanka was obtained from the Database of Global Administrative Areas (https://gadm.org/download_country36.html) and is freely available for academic use.
Fig 2.
Establishment of the serotyping method for LNKV and ANJZV infection based on the antigenic analysis of rGPs.
A: Antigenic profiling of rGPs of LNKV and ANJZV. The epitopes are mentioned as described by Arikawa et. al., [29]. B: Schematic diagram representing the arrangement of Gn and Gc of two virus GPs used in this study. C: IFA profiles of sera from host rodents at the serum dilution 1:1600. † Anti-LNKV serum from genome-positive and IgG-positive Mus booduga #PR106. § Anti-ANJZV serum from genome-positive and IgG-positive Rattus rattus #PR108. D: IFA titers and cross-neutralization titers of rodent sera used in panel C. As antibody negatives, sera from IgG-negative and genome-negative rodents, Mus booduga #PR99 ‡ and Rattus rattus #PR107 ‡ were used. 1Recombinant IFA antigens were transiently expressed in BHK/T7-9 cells and used for the endpoint titer determination. 2 Methods related to the alternative neutralization assay are shown in the S1 Text.
Table 1.
Hantavirus antibody positivity in febrile patients.
Fig 3.
Antibody titers in LNKV genome-positive febrile patient sera.
(A) Anti-LNKV antibody titers of P55 acute-serum and P100 acute and convalescent sera. (B) Serum antibody fluctuation graph of P100 at acute and convalescent phase sera. (C) Cross-reactivities of P55 and P100 to PUUV antigen. [35].
Fig 4.
Neighbor-Joining phylogenetic trees of partial nucleotide sequences.
(A) L-segment (347 bp: nt 2966-3312) (B) M-segment (218 bp: nt 2055-2272) and (C) S-segment (568 bp: nt 27-594), of newfound human hantaviruses and representative Muridae-borne hantaviruses. Representative Muridae-borne hantavirus sequences retrieved from databases were used to compare with newfound virus sequences (in boldface red) in this study. Previously identified Mus-borne LNKV sequences and rat-borne ANJZV sequences from Sri Lanka are in boldface. The scale bar indicates the sequence divergence values. The numbers above the nodes indicate the percentage consensus support values. The accession numbers of the sequences retrieved from the databases are mentioned in the legend of S1 Fig in the Supporting file.
Table 2.
Details of retrospective serological test results for patient sera with acute fever of unknown origin.
Table 3.
Details of the sample collections, geographical locations, and serological test results of CKDu patient and healthy control serum samples.