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Primers, kits and detection methods for detection of citrus Huanglongbing Asian species

A technology of citrus huanglongbing and detection kit, which is applied in the field of molecular genetic identification of molecular biology, can solve the problems of difficult separation and identification of amplified products, and achieves better specific detection effect, low possibility of false positives, and low probability of false positives. The effect of broad market prospects

Active Publication Date: 2019-03-19
广西特色作物研究院
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Problems solved by technology

However, the loop-mediated isothermal amplification technology has certain defects because the amplification product is a series of nucleic acid fragments ranging from large to small without identification of one or a few nucleic acid fragments of a specific length: (1) Amplified products are not easy to separate; (2) If there is non-specific amplification, it is difficult to distinguish and prone to false positives [6][9][10]

Method used

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  • Primers, kits and detection methods for detection of citrus Huanglongbing Asian species
  • Primers, kits and detection methods for detection of citrus Huanglongbing Asian species
  • Primers, kits and detection methods for detection of citrus Huanglongbing Asian species

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[0041] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0042] 1. Principle explanation

[0043] Such as figure 1 As shown, on the genome of H. citriensis Asiatic species, there are two upstream primer binding sites, which are two tandem repeat sequence units, and only one downstream primer binding site, which is a sequence-specific nucleic acid region. Since the DNA polymerase (SD enzyme) used in the reaction has dual characteristics of heat resistance and strand displacement activity, in the first round of reaction, the target nucleic acid fragment (double-stranded DNA) on the genome of Huanglongbing citri Denatured, unzipped into two DNA single strands, and then at 62°C annealing / extension temperature: the two upstream primers simultaneously bind to the DNA single-st...

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Abstract

The invention discloses a primer, a kit and a detection method for Candidatus Liberibacter asiaticus detection and belongs to the technical fields of molecular biology and molecular genetic identification. The invention designs a primer shown in SEQ ID NO.1-2 for the Candidatus Liberibacter asiaticus detection, a polymerase chain replacement reaction kit used for detecting Candidatus Liberibacter asiaticus is developed, and qualitative detection is carried out on the Candidatus Liberibacter asiaticus by adopting a polymerase chain replacement reaction process. The primer, kit and detection method for the detection have the characteristics of simple operation, high sensitivity, strong specificity and the like, and accuracy rate of molecular diagnosis on Candidatus Liberobacter asiaticum positive citrus plants can be improved; and meanwhile, operation is simple, and market prospect is broad, thereby being applicable to large-scale application.

Description

technical field [0001] The invention relates to primers, a kit and a detection method for detection of citrus huanglongbing bacteria Asian species, and belongs to the technical field of molecular biology molecular genetic identification. Background technique [0002] Huanglongbing (huanglongbing) is a worldwide destructive bacterial disease of citrus, and its pathogen is phloem-restricted hard-to-culture bacteria. So far, three species have been found: Candidatus Liberibacterasiaticus, Candidatus Liberibacter africanus and Candidatus Liberibacter americanus. Among them, the Asian species is the most widespread, and it is also the Asian species that is popular in my country. The economic loss caused by Huanglongbing to my country's citrus industry is as high as several billion yuan every year. Timely diagnosis and removal of Huanglongbing-infected trees in the field is an important part of the comprehensive prevention and control of Huanglongbing. Therefore, it is extremely...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2531/119C12Q2565/125
Inventor 娄兵海宋雅琴邓崇岭
Owner 广西特色作物研究院
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