Development of a reverse transcription-droplet digital PCR method for absolute quantification of citrus tatter leaf virus

被引：2

作者：

Zhao, Jinfa ^{[1
]}

Wang, Ying ^{[1
]}

Zhang, Xingkai ^{[1
]}

Zeng, Ting ^{[1
]}

Qin, Yangyang ^{[1
]}

Shen, Wanxia ^{[1
]}

Zhang, Wei ^{[1
,2
]}

Zhou, Yan ^{[1
]}

机构：

[1] Southwest Univ, Citrus Res Inst, Natl Citrus Engn & Technol Res Ctr, Chongqing 400712, Peoples R China

[2] Agr & Rural Affairs Sichuan Prov, Plant Protect Stn Dept, Chengdu 640041, Peoples R China

来源：

EUROPEAN JOURNAL OF PLANT PATHOLOGY | 2021年 / 161卷 / 03期

基金：

国家重点研发计划;

关键词：

citrus tatter leaf virus; Droplet digital PCR; Quantitation; ASSAY; APPLE;

D O I：

10.1007/s10658-021-02357-x

中图分类号：

S3 [农学（农艺学）];

学科分类号：

0901 ;

摘要：

Tatter leaf disease caused by citrus tatter leaf virus (CTLV) is a serious threat to the citrus industry. In this study, a reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR) assay for detection of CTLV was established by optimizing the reaction conditions. The dynamic range of the detection of CTLV RNA transcripts by RT-ddPCR was from 5 to 5 x 10(5) copies/mu L, and the sensitivity was 10 times higher than that of RT real-time PCR. Both techniques showed a high degree of linearity and quantitative correlation. The number of positive samples that was identified in field citrus samples using RT-ddPCR (62.3%) was higher than that of RT-qPCR (49.2%). The results demonstrated that the RT-ddPCR assay is a promising tool for improving CTLV detection.

引用

页码：743 / 749

页数：7

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