Abstract
BackgroundSchmallenberg virus (SBV), Akabane virus (AKAV) and Aino virus (AINV) are members of the Simbu serogroup within the genus Orthobunyavirus, family Bunyaviridae, which can cause reproductive disorders including abortion, stillbirth and congenital malformation in ruminants. Because, the clinical signs are similar, confirmatory diagnosis requires viral detection to differentiate infection between these three viruses.MethodsIn this study, a one-step multiplex reverse-transcriptase quantitative PCR (one-step mRT-qPCR) was developed for the simultaneous detection and differentiation of SBV, AKAV and AINV.ResultsThe detection limit of the one-step mRT-qPCR for SBV, AKAV and AINV were 2.4 copies (10 0.6 TCID 50/ml), 96.2 copies (10 1.5 TCID 50/ml) and 52.3 copies (10 1.2 TCID 50/ml), respectively. Various field samples such as bovine serum, bovine whole blood, bovine brain, goat serum and Culicoides were analyzed using the one-step mRT-qPCR and compared with previously published RT-qPCRs. The test results of the field samples were identical for the one-step mRT-qPCR and RT-qPCRs, which showed all samples to be negative for SBV, AKAV and AINV, except for one bovine brain sample (1/123) that was positive for AKAV.ConclusionThe one-step mRT-qPCR allows for the simultaneous detection of three viral pathogens (SBV, AKAV and AINV) that cause reproductive failure.
Highlights
Schmallenberg virus (SBV), Akabane virus (AKAV) and Aino virus (AINV) are members of the Simbu serogroup within the genus Orthobunyavirus, family Bunyaviridae, which can cause reproductive disorders including abortion, stillbirth and congenital malformation in ruminants
The Cp values and detection limit of the one-step mRT-qPCR were compared with the one-step single reverse-transcriptase quantitative PCR format for each of the viruses using the same templates
There were no appreciable differences in the mean Cp values between the one-step sRT-qPCR and one-step mRT-qPCR (Fig. 1)
Summary
Schmallenberg virus (SBV), Akabane virus (AKAV) and Aino virus (AINV) are members of the Simbu serogroup within the genus Orthobunyavirus, family Bunyaviridae, which can cause reproductive disorders including abortion, stillbirth and congenital malformation in ruminants. Since August 2011, a novel Orthobunyavirus named the Schmallenberg virus (SBV), which is closely related to simbu serogroup viruses, has been reported in ruminants such as cattle and sheep across Europe [8, 9]. These three bunyaviruses, SBV, AKAV and AINV are primarily transmitted by biting. The test was applied to clinical samples collected from ROK such as bovine serum, bovine whole blood, bovine brain, goat serum and Culicoides biting midges for further evaluation
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