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Development of a sensitive and reliable reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR) assay for the detection of Citrus tristeza virus
European Journal of Plant Pathology ( IF 1.7 ) Pub Date : 2020-01-13 , DOI: 10.1007/s10658-019-01920-x
Yingli Wang , Zhen Yang , Jinfa Zhao , Ruhui Li , Qin Wang , Jifen Li , Zhengwen Li , Yan Zhou

Citrus tristeza virus (CTV) is one of the most important citrus viruses in the world. In this study, we established a reverse-transcription droplet digital polymerase chain reaction (RT-ddPCR) method for the sensitive and accurate quantification of CTV. Quantitative linearity, sensitivity and accuracy of RT-ddPCR were compared to those of reverse-transcription real time PCR (RT-qPCR) by using 10-fold serial dilutions of the CTV RNA transcripts. Both methods showed a high degree of linearity (R2 = 0.991) and quantitative correlation, although RT-ddPCR revealed 100-fold higher sensitivity than RT-qPCR. The detection results for heat-treatment citrus samples also showed that the positive detection rate of RT-ddPCR (73.2%) was higher than that of RT-qPCR (53.6%). In summary, the results indicated that RT-ddPCR may contribute to improved analytical sensitivity and accuracy for CTV detection.



中文翻译:

灵敏可靠的反转录液滴数字聚合酶链反应(RT-ddPCR)检测试剂盒的开发,用于检测柑橘柑橘

特里斯柑橘病毒(CTV)是世界上最重要的柑橘病毒之一。在这项研究中,我们建立了逆转录液滴数字聚合酶链反应(RT-ddPCR)方法,用于对CTV进行灵敏而准确的定量。通过使用CTV RNA转录本的10倍系列稀释液,将RT-ddPCR的定量线性,灵敏度和准确性与逆转录实时PCR(RT-qPCR)进行了比较。两种方法均显示出高度的线性度(R 2 = 0.991)和定量相关性,尽管RT-ddPCR的灵敏度比RT-qPCR高100倍。热处理柑橘样品的检测结果还表明,RT-ddPCR的阳性检出率为73.2%,高于RT-qPCR的检出率为53.6%。总之,结果表明RT-ddPCR可能有助于提高CTV检测的分析灵敏度和准确性。

更新日期:2020-04-18
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