జర్నల్ ఆఫ్ ప్లాంట్ పాథాలజీ & మైక్రోబయాలజీ

నైరూప్య

Development of a Real-time RT-PCR Method for Rapid Detection and Quantification of Southern Rice Black-streaked Dwarf Virus in Rice

Songbai Zhang, Deyong Zhang, Yong Liu, Xiangwen Luo, Jue Cheng and Jing Peng

Southern Rice Black-Streaked Dwarf Virus (SRBSDV) was a new pathogen species in genus Fijivirus group 2, which caused serious yield loss of many important cereal crops, such as rice and maize, in the past four years (2008-2011), which are imported in China and Southeast Asia. Notably, there are no typical symptoms in early infected rice, which can prevent serious yield loss. So, it is very important for detection of the early stage of infection of SRBSDV in rice. To this end, a real-time RT-PCR method was established and used to detect this virus in rice samples, and it was also compared with the conventional RT-PCR. The result showed that the real-time RT-PCR possessed high specificity and sensitivity for SRBSDV detecting, and the relationship between Ct values and copy number of SRBSDV was linear with a range of 4.04×10²-4.04×10⁷ copies/reaction, and the sensitivity reached 150 copies/reaction. The intra- and inter-assay variability was low. The real-time RT-PCR detected SRBSDV in 87.5%- 100% of field rice samples compared to 63.6%-100% by conventional RT-PCR. As a whole, the real-time RT-PCR is a valuably alternative method for detecting SRBSDV in rice.