Fluorescent quantitative PCR method for detecting potato leaf roll virus

Abstract

The invention relates to the technical field of biological detection, in particular to a fluorescent quantitative PCR method for detecting a potato leaf virus. The fluorescent quantitative PCR methodcomprises the following steps that 1, RNA extraction is performed, wherein the total RNA of potatoes is extracted; 2, reverse transcription is performed, the extracted total RNA is reversely transcribed into cDNA; 3, primer design is performed, wherein two pairs of primers for amplification of an internal reference gene and a PLRV gene are respectively designed; 4, fluorescent quantitative PCR isperformed, the reversely transcribed cDNA and the designed primers are prepared into a PCR reaction system, and real-time fluorescent quantitative PCR reaction is performed. By adopting the detectionmethod, it can be detected at the molecular level that the potato leaf virus exists in potato plants, the virus gene expression situation can be quantitatively analyzed, and the fluorescent quantitative PCR method has the advantages of high efficiency, strong specificity, good accuracy and repeatability, high sensitivity, high detection speed, low material consumption, low cost, quantitative detection and the like.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a fluorescent quantitative PCR method for detecting potato leafroll virus. Background technique [0002] Potato leafroll virus (PLRV) is classified as Luteovirus group (Luteovirus), is a member of the genus PLRV, and is one of the most important viruses in potato virus diseases that endanger potato production. The PLRV virion is spherical, equiaxed, and 24nm in diameter. It is a positive-strand RNA virus without poly A at the 3' end and a strictly conserved sequence of 5-20 bp at the 5' end. The virus is transmitted in seed potatoes, and is transmitted by aphids in a persistent manner in the field. It cannot be transmitted mechanically. The vascular bundle of the host plant is its main distribution site. The content of the virus in the host body is low, and it is difficult to purify in large quantities, so that people cannot conduct thorough molecular biology resear...

AI Technical Summary

Problems solved by technology

Compared with real-time fluorescent quantitative PCR technology, other methods have some shortcomings, such as relatively long time-consuming, large consumption of detection materials, low sensitivity, and inability to quantitatively detect viruses, etc.

Images