Specific and rapid detection of Lily symptomless virus and Arabis mosaic virus in lily by dual IC-RT-PCR.

摘要

Lily symptomless virus (LSV) and Arabis mosaic virus (ArMV) cause severe losses of quantity and quality of lily flower and bulb production. Specificity, sensitivity and speed of detection methods for viruses need to be improved greatly to prevent LSV and ArMV from spreading from infected lilies. A dual IC-RT-PCR procedure for detection was developed in which the antibodies of LSV and ArMV were mixed and the mixture used to coat the PCR tubes. The particles of the two viruses were captured by the respective antibodies. Interference by other RNA viruses in infected lily was eliminated in the RT-PCR. Also, an RNA extraction step was omitted. The dual IC-RT-PCR products of LSV and ArMV were 521 bp and 691 bp, respectively. The specificity of the method was validated; only LSV and ArMV of four viruses were detected by dual IC-RT-PCR. The sensitivity of the detection method is 1 mg leaf tissue and higher than DAS-ELISA due to enrichment by dual immunocapture.