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Citrus yellow shoot candidatus liberibacter asiaticus detection test paper

A citrus huanglongbing detection test paper technology, which is applied in the field of citrus huanglongbing Asian pathogen detection test paper, can solve the problems that it is difficult to meet the needs of citrus orchard huanglongbing monitoring, and achieve the effect of low production cost, good stability and simple operation

Inactive Publication Date: 2013-01-09
INST OF PLANT PROTECTION FAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are difficult to meet the needs of monitoring Huanglongbing in citrus orchards

Method used

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  • Citrus yellow shoot candidatus liberibacter asiaticus detection test paper
  • Citrus yellow shoot candidatus liberibacter asiaticus detection test paper
  • Citrus yellow shoot candidatus liberibacter asiaticus detection test paper

Examples

Experimental program
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Effect test

preparation example Construction

[0032] (2) Preparation of antiserum

[0033] Experimental animals: healthy male white rabbits, each weighing more than 2kg.

[0034] Antigen emulsification: The emulsifiers are Freund's complete adjuvant and incomplete adjuvant, both purchased from SIGMA. Purified pathogenic outer membrane protein (prepared in step (1)), add an equal volume of Freund's complete or incomplete adjuvant, and pipette continuously until a drop of the mixed solution is not easy to spread on the surface of clear water.

[0035] Immunization: Before the first immunization, a small amount of blood was taken from the ear vein to prepare a healthy serum control. Rabbits were immunized with the above emulsified antigen five times, the first time was emulsified with Freund's complete adjuvant, and the last four times were emulsified with Freund's incomplete adjuvant; muscle multipoint method was used for immunization, with intervals of 7 days, 7 days, and 7 days in sequence. On d and 7 d, blood was colle...

Embodiment 1

[0045] Example 1 Detection of Periwinkle Samples with Huanglongbing Asian Species Pathogen

[0046] 1 material

[0047] Take 3-5 leaves with obvious symptoms from the periwinkle diseased plant with citrus huanglongbing Asian species pathogen, cut off the veins, grind them with distilled water, and the weight ratio of the veins to water is about 1:10. Then, let it sit for about 20 minutes.

[0048] 1.2 Method

[0049] Take the citrus huanglongbing pathogen detection test paper, insert one end of the sample pad into the sample solution to be tested for more than 0.5cm, but cannot submerge the colloidal gold pad, take out the sample liquid after swimming to the absorbent paper end for about 20 minutes, and then lay it flat and a dry surface of a level object, let stand for about 2 hours.

[0050] 1.3 Results and diagnosis

[0051] Such as figure 2 As shown, the sample with 2 red bands contains the Asian species of citrus Huanglongbing.

Embodiment 2

[0052] Example 2 Detection of Citrus Samples Carrying Huanglongbing Asian Species Pathogen

[0053] 1 material

[0054] Take 3-5 leaves with obvious symptoms from the diseased citrus citrus huanglongbing Asian species pathogen, cut off the veins, grind them with distilled water, and the weight ratio of the veins to water is about 1:10. Then, let it sit for about 20 minutes.

[0055] 1.2 Method

[0056] Take the citrus huanglongbing pathogen detection test paper, insert one end of the sample pad into the sample solution to be tested for about 0.5cm, take out the sample solution after swimming to the end of the absorbent paper for about 20 minutes, and then place it flat on the surface of a clean and dry horizontal object, Let stand for about 2 hours.

[0057] 1.3 Results and diagnosis such as image 3 As shown, there are 2 red bands, indicating that the Asian species of citrus greening disease is contained.

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Abstract

The invention belongs to the plant protection subject of the agricultural category, and particularly relates to citrus yellow shoot candidatus liberibacter asiaticus detection test paper. The detection test paper is prepared through the following steps that pathogenic bacteria antigen is extracted and purifed from catharanthus roseus host with citrus yellow shoot candidatus liberibacter asiaticus antigen and used for injecting immunized rabbits to obtain yellow shoot candidatus liberibacter asiaticus specific antiserum, the yellow shoot candidatus liberibacter asiaticus specific antiserum is purified to obtain the citrus yellow shoot candidatus liberibacter asiaticus antigen, immune colloidal gold is prepared, a combined pad which is soaked with the immune colloidal gold and a chromatography membrane which is coated with a detection line T-citrus yellow shoot candidatus liberibacter asiaticus antigen and a quality control line C-goat anti-rabbit antibody are further prepared, and finally a detection test paper product is assembled. Against for the requirement of the detection of a great amount of citrus yellow shoot samples, the citrus yellow shoot candidatus liberibacter asiaticus detection test paper which is convenient, rapid and high in specificity is provided. The test paper has following advantages that 1, convenience in use and simpleness in operation are realized; 2, a detection result can be obtained within a short period of time; 3, the stability is good; and 4, the cost is low, and the test paper is particularly suitable for the discrimination of citrus yellow shoot plants of a fruit garden.

Description

technical field [0001] The invention belongs to the field of agronomy and the subject of plant protection, and in particular relates to a test paper for detecting the pathogen of citrus huanglongbing Asian species. Background technique [0002] Citrus greening disease is a devastating disease in citrus production in the world, and it is listed as a quarantine disease by various countries. The causative agent of the disease is a phloem-limited endophytic bacterium called Cadidatus Liberibacter sp. Citrus huanglongbing is known to have Asian species ( Candidatus Liberibacter assiaticus ), African species ( Candidatus Liberibacter africanus ) and American species ( Candidatus Liberibacter amerinus ) three species. Huanglongbing in China is caused by an Asian pathogen. [0003] Citrus greening can be transmitted by scions, seedlings and vector psyllids (Asian psyllid, Diaphorina citri, African psyllid, Trioza erytreae). Huanglongbing pathogenic bacteria have many kinds o...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558
Inventor 刘利华朱祥枝陈军甘云陈玉妹
Owner INST OF PLANT PROTECTION FAAS
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