16Sr II group phytoplasmas PCR testing kit and testing process

Abstract

This invention relates to mycoplasma PCR detecting kit and its detecting method, it is specially used in 16SrII group mycoplasma detection. The kit includes PCR typic reaction reagent, mainly includes PCR reaction buffer solution, MgCl<2>,dNTPs,Taq DNA polyase, mycoplasma 16SrRNA gene general using primer and special primer of 16SrII group mycoplasma. The detection method includes PCR reaction and electrophoresis detection. 1500bp and 990bp two strip differential amplification stripes can be detected of the sample with 16SrII group mycoplasma, but there is only one 1500bp strip can be detected in the sample without 16SrII group mycoplasma. The using of this kit is convenient, and it has high specificity and detection sensitivity. It can directly detect mycoplasma disease sample and also can determine the mycoplasma that belongs to 16SrII.

Description

technical field [0001] The invention relates to an implant PCR detection kit and a detection method thereof, belonging to the field of biotechnology. Background technique [0002] Phytoplasma (formerly known as Mycoplasma-like Organism, MLO for short) is a kind of prokaryotic organism without cell wall that exists in plant sieve tube cells. Since phytoplasma was first discovered by Japanese scholar Toi Yoji in 1967, more than 300 plant phytoplasma diseases have been reported in the world so far. Because phytoplasma is strictly parasitic on the phloem of the host plant and cannot be cultured artificially, it is impossible to know its genetic nature, biochemical characteristics and nutritional requirements, and the development of its detection and identification technology is also hindered. The early identification of phytoplasma was mainly carried out through biological characteristics, such as symptom characteristics, and the relationship with insect mediators. These metho...

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Problems solved by technology

[0004] According to the phytoplasma 16SrRNA gene sequence, many specific primers for phytoplasma groups have been designed, including 16SrI group, 16SrIII group, 16SrV group, 16SrVI group, 16SrVII group, 16SrX group, etc. The 16SrRNA gene conserved region of the phytoplasma members of the 16SrII group is designed. Obviously, the specific primers cannot directly identify the phytoplasma of the 16SrII group.

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