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Primer, probe and detection method for real-time fluorescent polymerase chain reaction (PCR) of Planococcus lilacinus

A real-time fluorescence, mealybug technology, applied in the field of probes and detection, and real-time fluorescent PCR primers, can solve the problems of long time-consuming and difficult identification of scale insect morphology, and achieve long-term detection, rapid detection, and detection sensitivity. low effect

Inactive Publication Date: 2011-08-31
SHENZHEN AUDAQUE DATA TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide real-time fluorescent PCR primers, probes and detection methods for Pyeacoccus chinensis, aiming to solve the problems of time-consuming and difficult identification of scale insect morphology

Method used

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  • Primer, probe and detection method for real-time fluorescent polymerase chain reaction (PCR) of Planococcus lilacinus
  • Primer, probe and detection method for real-time fluorescent polymerase chain reaction (PCR) of Planococcus lilacinus

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Embodiment 1

[0033] Example 1 Preparation of primers and probes

[0034] Sequence determination and comparative analysis of the transcribed spacer (ITS) of P. chinensis and its close species, the sequences of the primers designed for the real-time fluorescent PCR detection of P. chinensis are as follows:

[0035] COI-PL-F: 5'-TAATGAAAATGTGCTACAACAAAATAAGTG-3';

[0036] COI-PL-R: 5'-AATTTTTAGATGAATAATAACCTTAAATGGAA-3'.

[0037] The sequence of the MGB probe used for the real-time fluorescent PCR detection of P. chinensis is as follows:

[0038] COI-PL-MGB: TTAGATAAAATGATACCTGTTAATC-MGB.

[0039] Among them, the fluorescent dye FAM is used as the reporter fluorescent group at the 5' end of the MGB probe, and the non-fluorescent quencher group (Non-Fluorescent Quencher, NFQ) is used as the quenching group at the 3' end, which does not generate fluorescence itself and can greatly reduce the The intensity of the background signal, and the 3' end of the probe is also connected with a MGB (Min...

Embodiment 2

[0041] The mealybug intercepted from imported Thai durian Durio zibethinus Murray was identified by real-time fluorescent PCR detection method with TaqMan MGB probe.

[0042] The primers and MGB probe used were those in Example 1.

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Abstract

The invention discloses a primer, a probe and a detection method for a real-time fluorescent polymerase chain reaction (PCR) of Planococcus lilacinus. The primer and the oligonucleotide probe which are applicable to the rapid detection of the real-time fluorescent PCR of the Planococcus lilacinus are designed, the disadvantages that the detection period is long, a specimen is required to be produced and rich experience is needed existing in the conventional morphological identification method are overcome, and the disadvantages that the operation is complicated or the detection sensitivity, the specificity, and the repeatability are low and the like existing in the conventional molecular detection method are also overcome; and the invention is particularly suitable for occasions with higher timeliness requirements such as port quarantine and the like because of a high detection speed and high specificity.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to real-time fluorescent PCR primers, probes and detection methods of Phenyococcus chinensis. Background technique [0002] Meainococcus argentina Planococcus lilacinus (Cockerell) is a pest in the "List of Entry and Exit Plant Quarantine Pests of the People's Republic of China", and it is also a quarantine pest that is of great concern to many countries or regions such as Europe and America. Pyracoccus chinensis is an important pest with a wide distribution in Southeast Asia and serious damage. There are many host plants of this insect, and it has been recorded that there are more than 100 species of host plants in 35 families, and it does damage to various fruits, trees and ornamental plants. The main host plants are durian, kumquat, mango, guava, custard apple, cherries, longan, rambutan, pomegranate, betel nut, jackfruit, erythrina, cocoa, sheep hoof, plantain, peach oliv...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 焦懿徐浪余道坚陈志粦康林
Owner SHENZHEN AUDAQUE DATA TECH