Detection of IgM and IgG in the Sera Samples from Patients with Unspecified Fever Collected in Republic of Guinea, Using the Developed Microarray Based on the Recombinant Antigens of Arboviruses (ZIKV, DV, CHIKV, CCFV, RVFV)

Background: The highly informative screening method of the indirect serological diagnostics, e.g. antibody detection, is needed during arboviruses-caused infection because of short viremia period and non-specificity of the first clinical signs for most these diseases. Here we aimed to develop and evaluate the recombinant protein based microarray for detection of IgM/IgG to antigens of Dengue, Zika, Chikungunya, Сrimean-Congo and Rift-Valley fever viruses. Methods & Materials: The microarray development stage was conducted using sera from Russian patients in which DV, ZIKV, CCFV RNA have been determined using the RT-PCR systems (CRIE, Russia) (122 in total). The evaluation stage was conducted using negative for anti-YFV IgM sera from Guinean patients with unspecified fever (163 in total). Results: The developed microarray includes recombinant NS1 and E antigens for DV and ZIKV; E1 and E2 – for CHIKV; G1, G2, Np for CCFV and RVFV. In 9/9 ZIKV-cases IgM were discovered in sera on 3-5 day and IgG on 5-8 day after symptoms onset. In 43/60 PCR-positive DV-cases IgM to DV type 1 or type 3 antigens were discovered and in 22/60 – IgG. 3/4 CHIKV PCR-positive cases were confirmed by IgM detection and ¼ - by IgG detection in sera. 17/20 CCFV-cases were confirmed by detection of specific IgM or IgG in early samples and 20/20 – in late samples. IgM/IgG to all recombinant antigens weren’t found in reference cohort (50 samples) under assay conditions. From 163 Guinean samples in absence of IgG IgM to arboviruses antigens were found in 16 sera (9.8%), indicating about acute infection stage. From that 16 cases in 4 cases anti-ZIKV IgM were detected, in 2 cases – anti-DV, in 2 cases – to unspecified flavivirus, in 1 case – anti-CHIKV, 4 cases – anti-CCFV, in 2 cases – anti-RVFV. Conclusion: We showed that developed microarray is highly sensitive and specific diagnostic mean. When IgM response to arboviruses antigens is investigated the usage of the technique could be useful for rapid diagnostics as it could help to combine several ELISAs investigations in one test. So the developed microarray could be considered as time- and resource-saving perspective screening test.