Real-time fluorescence isothermal-amplification primer groups and reaction system and detection method for quantitatively detecting locus rot germ

Abstract

The invention discloses a set of real-time fluorescence isothermal-amplification (Real-time LAMP) primer groups designed and obtained according to locus-rot-germ mitochondria small subunit (mtSSU rRNA genes) gene sequences. The set of real-time fluorescence isothermal-amplification (Real-time LAMP) primer groups comprises a pair of outer primers F3 and B3 and a pair of inner primers FIP and BIP, and a real-time fluorescence loop-mediated isothermal amplification detection system of the locus rot germ is set. The real-time fluorescence isothermal-amplification reaction system comprises the set of real-time fluorescence isothermal-amplification primer groups, fluorescent dyes, reaction liquid and Bst DNA polymerase, to-be-detected sample DNA is added, the fluorescence intensity of the reaction system is detected in real time along with an isothermal-amplification reaction, the Ct value is calculated according to analysis software on a real-time fluorescence analyzer, and therefore the DNA content of a locus rot germ in to-be-detected DNA extracting liquid is calculated. The method is easy and convenient to operate, rapid, good in specificity and high in sensitivity, the locus rot germ can be rapidly and quantitatively detected, and the condition that plants are infected by the locus rot germ can be really responded; the detection method is suitable for widely applied and popularized in the plant-disease diagnosis detection field.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a real-time fluorescent isothermal amplification (Real-time LAMP) primer set, a reaction system and a detection method for quantitative detection of lotus spoilage bacteria in soil and plants. Background technique [0002] Lotus rot is caused by Fusarium commune, which often causes the whip, joints, and roots of the lotus to rot. At the same time, the leaves and flowers on the ground will also wilt and yellow, and the whole plant will die if the disease is severe. The lotus is widely planted, and lotus rot is common and harmful. It has become a major obstacle restricting the development of the lotus industry in Guangdong Province and even the whole country. It is also an important bottleneck problem in the development of urban agriculture and greening of wetlands in my country. [0003] In recent years, it has been reported that the PCR-based method has been successfully ...

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Problems solved by technology

[0003] In recent years, it has been reported that the PCR-based method has been successfully used in the detection of lotus spoilage bacteria, but the public detection methods are all qualitative detection, and there is no report on the rapid quantitative detection method of lotus spoilage bacteria

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