Abstract:

To develop a rapid and reliable detection method，a real-time RT-PCR system based on SYBR GreenⅠwas established by using a pair of specific primers designed from conserved coat protein gene of Citrus yellow vein clearing virus（CYVCV）. The sensitivity，specificity and applicability of the system were evaluated accordingly. The results showed that amplicons were produced from CYVCV isolates，whereas no amplicons from non-CYVCV citrus virus samples including Citrus tristeza virus （CTV）and Citrus tatter leaf virus（CTLV）were obtained. The sensitivity of the real-time RT-PCR was 100-fold higher than that of conventional RT-PCR. An excellent linear correlation（R2 = 0.999）obtained from two standard curves of cRNA and the amplification efficiency was 102%. The data from field citrus samples detection showed that the real-time RT-PCR system could be used to determine the concentration of CYVCV in the different citrus species.

Key words: citrus, Citrus yellow vein clearing virus, real-time RT-PCR, detection, application