玉米褪绿斑驳病毒RT-PCR检测引物的筛选

Screening of primers for RT-PCR detection of maize chlorotic mottle virus

收集18对已报道的玉米褪绿斑驳病毒(MCMV)RT-PCR检测引物,利用MCMV阳性样品和健康样品进行测试,对实际表现较优的引物进一步分析其特异性和灵敏度,再以220份玉米种子样品进行验证。结果显示,在测试的18对引物中,有2对引物MCMV PF7/MCMV PR7和M-PCR-F/M-PCR-R对MCMV的检测效果较优,其对7种其他玉米病毒均无交叉反应,检测灵敏度可达到200 pg/μL;实际样品检测中,应用上述2对引物检测的结果一致,均检出6份阳性样品。根据测试结果,推荐使用引物MCMV PF7/MCMV PR7和M-PCR-F/M-PCR-R用于玉米种子携带MCMV的检测。

To evaluate the reported specific primers for the RT-PCR detection of maize chlorotic mottle virus(MCMV),we tested both MCMV positive and negative samples with 18 pairs of primers collected in this study.The specificity and sensitivity of the selected primers were further analyzed,and then 220 maize samples were used for verification.The results indicated that among the 18 pairs of primers,two pairs of primers MCMV PF7/MCMV PR7 and M-PCR-F/M-PCRR had superior detection performance for MCMV.There was no cross-reaction with seven other main viruses of maize.The detection sensitivity of the selected primers could reach 200 pg/μL.Out of the 220 maize samples,six were determined as MCMV-positive by using the primers MCMV PF7/MCMV PR7 and M-PCR-F/M-PCR-R.Based on these findings,the two pairs of reported primers,MCMV PF7/MCMV PR7 and M-PCR-F/M-PCR-R,were recommended for the detection of MCMV in maize seeds.