Real time TaqMan RT-PCR assay for the detection of Cucumber green mottle mosaic virus.
A real time reverse-transcription polymerase chain reaction (RT-PCR) was developed for efficient detection of Cucumber green mottle mosaic virus (CGMMV). The method was designed to use a duo-primer system with a TaqMan probe targeting the conserved sequence in 3' noncoding region (NCR) of CGMMV to detect isolates of this virus collected in China. The sensitivity of the real time RT-PCR assay was 0.13 pg of total RNA or 50 molecules of RNA transcripts. This level of sensitivity indicated that the one step real time RT-PCR developed in the present study could be used for routine testing assays. The real time RT-PCR method could assist in the implementation of quarantine measures for prevention and control of the disease caused by CGMMV.
Hongyun
Chen
C
Xiamen Entry-Exit Inspection and Quarantine Bureau, Xiamen 361012, China.
Wenjun
Zhao
Z
Qinsheng
Gu
G
Qing
Chen
C
Shiming
Lin
L
Shuifang
Zhu
Z
D020413:3' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D002681:China; D018553:Cucumis sativus / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017898:Tobamovirus / Q000235:genetics / Q000302:isolation & purification*
D020413:3' Untranslated Regions; D017931:DNA Primers; D012367:RNA, Viral