Detection of Tomato spotted wilt virus in its vector Frankliniella occidentalis by reverse transcription-polymerase chain reaction.

A method for rapid and reliable detection of Tomato spotted wilt virus (TSWV) (Tospovirus, Bunyaviridae) in its vector Frankliniella occidentalis (Thysanoptera Thripidae) would be a useful tool for studying the epidemiology of this virus. A RT-PCR method developed for this purpose is reported. The method was tested on thrips involved in laboratory transmission trials and on thrips collected in the field, whose capability to transmit TSWV was checked previously by leaf disk assays. The RT-PCR results were consistent with the results obtained by the leaf disk assays. Among thrips involved in laboratory experiments, 97% of the adults that transmitted TSWV were positive by RT-PCR; as did some non-transmitter adults reacted, whereas among field-collected thrips only the individuals able to transmit were positive by RT-PCR. In addition, healthy thrips were allowed to feed as adults on virus-infected leaves for 48 h, and then examined by RT-PCR immediately or after starving or feeding on virus-free plants for various times, to determine if virus ingested (but not transmissible) was also detectable. The virus was detectable immediately after the feed or within 12 and 24 h for individuals starved or fed on virus-free plants, respectively, but not after those periods. Thus, the method could detect rapidly and reliably the virus in vectors from the field, providing 24 h of starving to avoid positive RT-PCR results from thrips simply carrying the virus.