免疫磁珠结合RT-PCR检测黄瓜绿斑驳花叶病毒

Detection of cucumber green mottle mosaic virus by immunomagnetic separation and RT-PCR

以感染黄瓜绿斑驳花叶病毒的黄瓜种子为材料,建立了免疫磁珠结合RT-PCR直接检测种子携带病毒的高灵敏度方法。结果表明:当磁珠的包被单克隆抗体浓度为1.5×10^(-5)mg/mL时,可检测到的最低抗原浓度为7.813×10^(-3)mg/mL,相当于8 ng种子中的黄瓜绿斑驳花叶病毒。该方法的灵敏度比免疫捕获RT-PCR(IC-RT-PCR)的灵敏度高出8倍。

The cucumber seeds infected with cucumber green mottle mosaic virus were used as a test material,and a high sensitivity method of directly detecting the virus was established by immu-nomagnetic separation and reverse transcriptase-polymerase chain reaction（IMS-RT-PCR）.The test results showed that when the concentration of envelope monoclonal antibody of magnetic beads was 1.5×10^（-5） mg/mL,the lowest detectable antigen concentration was 7.813×10^（-3） mg/mL,about a quantity of the virus in 8 ng cucumber seeds,indicating the sensibility of this method was 8 times higher than that of the immunocapture RT-PCR（IC-RT-PCR）.