Specific primer for quantitatively detecting yellow leaf curl virus, as well as application of specific primer

Abstract

The invention belongs to the field of pathogen detection, and provides a primer pair for quantitatively detecting a yellow leaf curl virus, application of the primer pair to quantitative detection of the yellow leaf curl virus and a method for quantitatively detecting the yellow leaf curl virus. The primer pair for quantitatively detecting the yellow leaf curl virus can be used for efficiently and accurately amplifying a target fragment. The method for quantitatively detecting the yellow leaf curl virus by virtue of the primer pair can be used for accurately and rapidly identifying pathogenic bacteria and accurately quantifying a viable count.

Description

technical field [0001] The invention belongs to the field of pathogen detection, in particular to a specific primer for quantitative detection of yellow leaf curl virus and its application. Background technique [0002] Traditional detection methods of pathogenic bacteria, such as physiological and biochemical index identification methods, bacterial isolation and culture identification methods, etc., rely on phenotypes to identify pathogenic bacteria; these methods are time-consuming, have low sensitivity, and poor accuracy. [0003] Propidium azide bromide (PMA) is a fluorescent dye with high affinity to DNA, which can pass through the damaged cell membrane and covalently cross-link with DNA under the irradiation of visible light, so as to block the DNA in damaged cells. The role of DNA for PCR amplification. Some studies have reported that the PMA-qPCR method combined with the detection of live pathogen content, but only applied in water (Fujimoto J, Watanabe K. Quantitat...

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Problems solved by technology

Although the common qPCR method can quantify the pathogenic bacteria, it cannot distinguish between the dead and the dead of the pathogenic bacteria (Zhang Zhenjia, Yu Jihua, Wang Xilin. Preliminary report on the control test of bacterial angular spot of cucumber[J]. Journal of Gansu Agricultural University, 1989, 4 :63-66.; Tong Tiezheng, Wu Shuxu, Li Dan, et al. Study on the disinfection characteristics of pathogenic bacteria based on PMA-quantitative PCR selective detection technology[J].Environmental Science,2011,32(4):1120-1126.)

Studies have shown that the DNA of pathogenic bacteria can remain intact within days or even weeks after death (Josephson K, Gerba C, Pepper I. Polymerase chain reaction detection of nonviable bacterial pathogens [J]. Applied and Environmental Microbiology, 1993, 59 (10 ):3513-3515.), so how to distinguish the life and death of pathogens has become a huge challenge for molecular detection methods

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