Browsing by Subject "Puumala virus"

Puumala orthohantavirus (PUUV) is a single stranded negative sense RNA virus, carried by the bank vole (Myodes glareolus). Like other orthohantaviruses, it does not cause visible symptoms in the host species, but when transmitted to humans, it can cause a mild version of hemorrhagic fever with renal syndrome (HFRS) called nephropathia epidemica (NE). PUUV is the only pathogenic orthohantavirus that is endemic to Finland, where it has a relatively high incidence of approximately 35 in 100 000 inhabitants or 1000 to 3000 diagnosed cases annually. Here we describe a miniaturized immunofluorescence assay (mini-IFA) for measuring antibody response against PUUV from bank vole whole blood and heart samples as well as from patient serum samples. The method outline was based on the work done by Pietiäinen et al., (2022), but it was adapted for the detection of PUUV antibodies. Transfected cells expressing the PUUV structural proteins (N, GPC, Gn and Gc) were used instead of PUUV infected cells, which allowed for performing all steps outside of bio-safety level 3 (BSL3) conditions. This method also enables the simultaneous measurement of IgM, IgA and IgG antibody response from each sample in a more efficient and higher output manner, when compared to traditional immunofluorescence methods. Our results show that the method is effective for testing large amounts of samples for PUUV antibodies and it allows for quick and convenient access to high-quality images that can be used for both detecting interesting targets for future studies, as well as producing a visual archive of the test results.