Abstract
Alphaviruses have a single-stranded, positive-sense RNA genome that contains two open reading frames encoding either the non-structural or the structural genes. Upon infection, the genomic RNA is translated into the non-structural proteins (nsPs). NsPs are required for viral RNA replication and transcription driven from the subgenomic promoter (sgP). Transfection of an RNA encoding the luciferase gene under the control of the sgP into cells enabled the detection of replication-competent chikungunya virus (CHIKV) or Mayaro virus (MAYV) with high sensitivity as a function of the induced luciferase activity. This assay principle was additionally used to analyze virus-neutralizing antibodies in sera and might be an alternative to standard virus neutralization assays based on virus titration or the use of genetically modified tagged viruses.
Highlights
Alphaviruses are arthropod-transmitted viruses of the Togaviridae family
NsPs are required for viral RNA replication, which includes the synthesis of a minus-sense RNA from the genomic RNA (gRNA), resulting in double-stranded RNA intermediates
The minussense RNA serves as a template for the gRNA and the subgenomic RNA, the synthesis of which is controlled by the subgenomic promoter
Summary
Members of the Old World alphaviruses, such as chikungunya virus (CHIKV), Mayaro virus (MAYV), and Ross River virus (RRV), cause an acute infection with high fever, rash, myalgia, and polyarthralgia that lasts around 4–7 days, with viremia and high viral loads. It is only during this viremic phase that the virus can be detected in the blood by nucleic acid testing. The genomic RNA (gRNA) is directly translated into a non-structural precursor protein, which is processed by viral and cellular proteases. The structural proteins are translated from the sgRNA [3]
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