果树学报2011，Vol.28Issue(3)：458-462,5.
柑橘碎叶病毒巢式RT-PCR检测方法建立及应用
Establishment of nested RT-PCR for detecting Citrus tatter leaf virus
摘要
Abstract
Citrus tatter leafvirus (CTLV) is an economically important pathogen of citrus. To establish a sensitive and specific method for detection of this virus, a nested RT-PCR was developed by comparing the detection efficiency of five primer sets. Total nucleotide acid extraction of CTLV-inoculated samples were serially tenfold diluted to evaluate the sensitivity, and samples infected with CTV, SDV, HLB,Canker, or CEVd were used to determine the specificity. The results showed that the lowest detection limit of the total nucleic acid by the method reached about 1.27 μg· L-1, which was hundred times lower than that of One-step RT-PCR assay at least. No positive result was obtained from the samples without CTLV using this method. In the detection of 142 samples collected from orchards, 24 shoot-tip grafting citrus,and 20 rusk citrange infected by CTLV, the positive rates using the nested RT-PCR, the routine seminested PCR, and the one-step RT-PCR were 21.5％, 18.3％ and 19.4％ respectively. It suggested that the nested RT-PCR is a sensitive, specific method and can be used as a useful tool for CTLV detection.关键词
柑橘碎叶病毒/巢式RT-PCR/检测技术Key words
Citrus tatter leaf virus/ Nested RT-PCR/ Detection分类
农业科技引用本文复制引用
宋震,周常勇,刘科宏,李中安..柑橘碎叶病毒巢式RT-PCR检测方法建立及应用[J].果树学报,2011,28(3):458-462,5.基金项目
公益性行业科研专项(NYHYZX07-023,200903004-06) （NYHYZX07-023,200903004-06）
中央高校基本科研业务专项(XDJK2009C131) （XDJK2009C131）
重庆市自然科学基金(CSTC,2010BB115) （CSTC,2010BB115）
