The methods of NCM-ELISA, RT-PCR and semi-nested RT-PCR were used to detect the Sweet potato chlorotic stunt virus (SPCSV) in the whitefly Bemisia tabaci, and the sensitivities of these methods were compared. The results showed that SPCSV could be detected from at least 16 whiteflies carrying the virus by NCM-ELISA, occasionally from single whitefly and steadily from more than three whiteflies by RT-PCR; SPCSV could be steadily detected from single insect by semi-nested RT-PCR. With in vitro RNA transcript as template, the sensitivity were up to 5. 69 × 104 copies/FL and 5. 69 ×101 copies/μL by RT-PCR and semi-nested RT-PCR, respectively, indicating that the detection sensitivity of the semi-nested RT-PCR was 1 000 times higher than that of the RT-PCR.%建立了利用NCM-ELISA、RT PCR和半巢式RT-PCR检测烟粉虱中SPCSV的方法,比较了3种检测方法的灵敏性.结果表明,NCM-ELISA最低能从16头带毒烟粉虱中检测出SPCSV;RT-PCR能稳定地从3头以上带毒烟粉虱中检测出SPCSV;半巢式RT-PCR能稳定地从单头烟粉虱中检测出SPCSV.检测灵敏性研究表明,用体外转录的RNA作为核酸模板,RT-PCR可检测的最低浓度为5.69×104拷贝/μL,半巢式RT-PCR为5.69×101拷贝/μL,说明半巢式RT PCR检测方法的灵敏性比RT-PCR高1 000倍.
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