Kit and method for detecting plum pox virus by applying nano magnetic bead

A technology of nano-magnetic beads and plum pox virus, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection, which can solve the problems of easy false positive results, cumbersome RNA extraction, and low detection sensitivity, etc. question

Inactive Publication Date: 2016-07-13
INSPECTION AND QUARANTINE TECHNOLOGY CENTER ZHONGSHAN ENTRY EXIT INSPECTION AND QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional ELISA technology has low detection sensitivity, long detection time, and prone to false positive results in the actual application process; RT-PCR technology has high sensitivity, but the RNA extraction is cumbersome and easy to degrade, and the PCR reaction is easily affected by polyphenols in the host. , polysaccharide substances, it is difficult to realize the rapid, simple and efficient detection of plant virus disease samples

Method used

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  • Kit and method for detecting plum pox virus by applying nano magnetic bead
  • Kit and method for detecting plum pox virus by applying nano magnetic bead
  • Kit and method for detecting plum pox virus by applying nano magnetic bead

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Embodiment 1

[0063] The present invention uses nano-magnetic beads to detect the method for plum pox virus:

[0064] 1. Identification and storage of virus samples

[0065] Identification: The collected samples are first detected by serological methods, mainly ELISA, and confirmed by modern molecular biological methods such as PCR detection according to the serological test results. Preservation: select leaf specimens with typical symptoms, freeze and dry them quickly, and store them in a -80°C refrigerator;

[0066] 2. Preparation of magnetic beads

[0067] 1). Fe 3 o 4 Preparation and amination modification of magnetic nanoparticles (refer to the method of Beijing Jinna Technology Co., Ltd.)

[0068] The specific operation is: take 5.2g FeCl respectively 3 ·6H 2 O, 2.7g FeSO 4 ·7H 2 O. Dissolve 0.85mL of 12.1moL / L concentrated hydrochloric acid in 200mL of water, ultrasonically deoxygenate, and then drop the above solution into 250mL of 0.75moL / L NaOH solution. All reactions wer...

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Abstract

The invention discloses a kit and method for detecting plum pox virus by applying a nano magnetic bead. The kit contains primers PPV1, namely 5'-ACCGAGACCACTACACTCCC-3', and PPV2, namely 5'-CAGACTACAGCCTCGCCAGA-3'. The method for detecting the plum pox virus by applying the nano magnetic bead is characterized by comprising the following steps: 1) preparing a sample; 2) cleaning the nano magnetic bead; 3) combining; 4) cleaning; 5) carrying out splitting decomposition; 6) sampling; 7) carrying out RT-PCR amplification, and after reaction is finished, taking 5mu L reaction product for carrying out electrophoretic analysis on 1% agarose gel prepared by adopting 1*TAE buffer solution. The invention aims at overcoming the defects of the prior art and provides the kit and the simple method for detecting the plum pox virus by applying the nano magnetic bead.

Description

technical field [0001] The invention relates to a kit and a method for detecting plum pox virus using nanometer magnetic beads. Background technique [0002] Plumpox virus, PPV is one of the most dangerous viruses in drupe fruit trees. Once the fruit tree is infected by it, it will cause a large number of unripe fruits to fall off, severely reduce the yield, and greatly reduce the quality of the fruit. The speed of transmission is fast, and it may bring catastrophic losses to the development of the fruit tree industry of the whole country in a short period of time. The virus was first discovered in Bulgaria in 1915, and then quickly spread to most countries in Europe, as well as the Mediterranean, Central Africa, India and Chile, and has also been reported in the United States and Canada. Because of the seriousness of the virus, it has been listed as a quarantine pathogen of type A2 by the European Plant Protection Organization. A series of strict detection and eradication...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701C12Q1/6806
Inventor 陈定虎杨雷亮管维杨志方吴颖儿
Owner INSPECTION AND QUARANTINE TECHNOLOGY CENTER ZHONGSHAN ENTRY EXIT INSPECTION AND QUARANTINE
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