Primer group and application of primer group in rapid detection of leptosphaeria lindquistii Frezzi based on micro-fluidic chip

Abstract

The invention relates to a primer group and application of the primer group in rapid detection of leptosphaeria lindquistii Frezzi based on a micro-fluidic chip. The primer group is composed of the following primers: LI-F3-4, LI-B3-2 / 3 / 4, LI-FIP-4, LI-BIP-4, LI-LF-4 and LI-LB-4. The primer group disclosed by the invention is strong in amplification specificity and high in sensitivity, and the lowest detection limit can reach 101 copies / [mu]L. The detection method disclosed by the invention is high in detection speed, and can be used to simultaneously detect a variety of pathogens. The peak initiation time is 7-12 min, and the whole process consumes 0.5 h, so that the detection speed is substantially accelerated, and work efficiency is improved. Moreover, sensitivity is high, and the reaction system is small, so that cost is saved, no expensive instrument is required, operation is simple, and the primer group is suitable for rapid screening at ports and other non-laboratory places.

Description

technical field [0001] The invention relates to the detection technology of plant pathogenic fungi, in particular to a primer set and its application in rapid detection of sunflower black stem fungus based on a microfluidic chip. Background technique [0002] The sunflower black stem fungus is a devastating disease that sunflowers are very susceptible to during the growth process. In 2010, the pathogen was included in the "List of Imported Plant Quarantine Pests of the People's Republic of China", and it is my country's imported plant quarantine pests. [0003] At present, sunflower black stem disease is mostly distributed in countries and regions such as Canada, the former Yugoslavia, France, Argentina, Hungary, Romania, Australia, Iran, Pakistan and the United States. In 2008, it was first reported that the disease occurred in Xinjiang, and then it also occurred in Jilin, Inner Mongolia, and Hebei. In the early stage of sunflower black stem disease, black spots appear at...

AI Technical Summary

Problems solved by technology

Among them, the biological identification method is labor-intensive and time-consuming, and it is difficult to distinguish the sunflower black stem fungus from other Phomomycetes morphologically, so the identification is difficult

However, the hardware platform required for preliminary screening of samples by PCR method is relatively expensive, and it is not easy for grassroots testing institutions to popularize

At the same time, due to the low sensitivity of PCR with specific primers, DNA samples with lower content cannot be detected, and the detection accuracy cannot be met.

In addition, if gene sequence analysis is used, although the amount of biological information obtained is large, sequencing is required, and the detection cycle is long

Although RFLP labeling technology does not require sequencing, multiple enzyme digestions are also time-consuming and labor-intensive.

In addition, the above molecular detection methods must be subjected to electrophoresis, and the experimenters also need to be exposed to toxic reagents, and cross-contamination is prone to occur, resulting in false positive results

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