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Wheat streak mosaic virus (WSMV) is transmitted by the wheat curl mite (WCM), Aceria tosichella. Immunofluorescent and dot-immunobinding assays were developed to detect the presence of WSMV in single WCM. Virus-specific immunofluorescent microscopy detected WSMV near the anterior end of viruliferous WCM. With dot-immunobinding assay, WSMV was detected in WCM fed on WSMV-infected wheat (Triticum aestivum) but not in WCM maintained on healthy plants. Both immunofluorescent and dot-immunobinding assays were sufficiently sensitive to detect WSMV in individual WCM, providing a means to determine the percentage of viruliferous WCM in field collections.