| Popis: |
Rice is one of the main staple foodstuffs for the world's population; however, several diseases caused by phytopathogens cause yield losses in rice crop. Rice crinkle is an emerging disease in rice cultivation, caused by rice stripe necrosis virus (RSNV). RSNV is transmitted by the soil plasmodiophoromycete, Polymyxa graminis, which can remain in the soil through resistance structures for decades. The symptoms caused by the virus are often confused with other causes, making a correct diagnosis and the adoption of effective management and control measures difficult. RSNV is detected through tests such as plate trapped antigen – enzyme-linked immunosorbent assay (PTA-ELISA) and conventional reverse transcription polymerase chain reaction RT-PCR. The objective of this study was to develop a protocol for RSNV detection via real-time RT-PCR using a TaqMan probe associated with direct sample preparation methods, which has not been published to date. The RT-PCR detection protocol developed was 100–1,000 times more sensitive than conventional RT-PCR, allowing for the detection of RSNV in contaminated plant material and soil. The direct sample preparation method associated with real-time RT-PCR exhibited similar sensitivity to the nucleic acid purification methods. |
