Serologic detecting kit for potyvirus on sweet potato and detecting method thereof

A detection kit and detection method technology, applied in the field of bioengineering, can solve problems such as waste, time-consuming and labor-intensive, and achieve the effects of convenient detection, quality assurance, and cost-saving detection

Inactive Publication Date: 2012-06-27
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

due to infestation of sweet potato Potyviruses There are many types of viruses. When it is not necessary to determine the type of virus, it is only necessary to determine whether the sample contains Potyviruses Viruses (such as the detection of virus-free stem tip seedlings of sweet potato), when using the above method for detection, it is necessary to detect various viruses one by one, which is often time-consuming and laborious, resulting in waste

Method used

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  • Serologic detecting kit for potyvirus on sweet potato and detecting method thereof
  • Serologic detecting kit for potyvirus on sweet potato and detecting method thereof
  • Serologic detecting kit for potyvirus on sweet potato and detecting method thereof

Examples

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Effect test

Embodiment 1

[0033] Embodiment 1, Kit for the serological detection of Potavirus genus virus on sweet potatoes, see figure 1 , in the figure 1 is the upper layer of the kit, 2 is the reagent bottle, and 3 is the lower layer of the kit.

[0034] The kit contains four kinds of specific antibodies of potyvirus genus virus and a mixed antibody. The kit is divided into upper and lower layers, and a partition is arranged between the upper and lower layers. Reagent bottle jack, reagent bottles are placed in the jack, the lower layer is a push-pull drawer, and nitrocellulose membrane is placed in the drawer; there are nine reagent bottles on the upper layer, which are No. 1 to No. 9 reagent bottles, of which No. 1 Sweet potato feather mottle virus SPFMV antibody in the reagent bottle, No. 2 reagent bottle is sweet potato latent virus SPLV antibody, No. 3 reagent bottle is sweet potato G virus SPVG antibody, No. 4 reagent bottle is sweet potato vein mosaic virus SPVMV antibody, No. 5 The reagen...

Embodiment 2

[0037] Embodiment 2, The detection method of the serological detection kit of potyvirus genus virus on sweet potato, comprises the following steps:

[0038] 1. Preparation of buffer solution:

[0039] (1) TBS: 0.02M Tris+0.50M NaCl, pH7.5;

[0040](2) Extraction buffer: TBS+0.2% sodium sulfite;

[0041] (3) Blocking solution: TBS+2% skim milk powder+2% triton X-100;

[0042] (4) Antibody buffer: TBS+2% skimmed milk powder;

[0043] (5) T-TBS: TBS+0.05% Tween 20;

[0044] (6) Substrate buffer: 0.1M Tris+ 0.1M NaCl+5mM MgCl 2 . 6H 2 O, pH=9.5;

[0045] Where M refers to the unit mol / L.

[0046] 2. Detection steps:

[0047] (1) Prepare nitrocellulose membrane (NC membrane): Cut out an appropriate size of NC membrane and mark it with a pencil to facilitate sample application.

[0048] (2) Sample treatment: Quick-freeze 0.1g sweet potato sample in liquid nitrogen and grind it into powder, add 100μl extraction buffer (TBS+ 0.2% sodium sulfite) and continue grinding until...

Embodiment 3

[0061] Embodiment 3, sweet potato on Potyviruses Application of virus serological detection kit

[0062] Using this kit, 24 field sweet potato samples collected from the field were detected. The detection steps are as in Example 2. The primary antibody uses mixed antibody antiserum (No. 5 reagent bottle). The working concentration is 1:1000 times. The results showed that 10 of the 24 sweet potato samples were positive (see figure 2 ), the positive rate was 41.7%, indicating that these sweet potato samples are commonly found in Potyviruses Virus.

[0063] figure 2 Among them, the upper row 1, 2, 3, 4, 5, 6, 7, 8, and the lower row 1, 2 are positive samples.

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Abstract

The invention relates to a serologic detecting kit for potyvirus on a sweet potato and a detecting method thereof. The kit is divided into an upper layer and a lower layer, wherein reagent bottles are arranged on the upper layer; a push-pull drawer is arranged on the lower layer; a nitrocellulose film is placed into the drawer; and a specific antibody for the potyvirus, a mixed antibody for the potyvirus and a goat anti-rabbit IgG, NBT, BCIP and Tris-HCl solution marked with alkaline phospholipase are respectively filled in different reagent bottles. When a sample is detected, the reagent bottle containing the mixed reagent is used for detecting if the sample contains the potyvirus; other specific antibodies are used for detecting when a certain specific virus needs to be confirmed; the detection is convenient; and the cost is saved; and the detecting method is simple, direct, convenient and practical. The serologic detecting kit and the detecting method provided by the invention can be applied to the detection for the detoxified sweet potato stem seedling and all-level potatoes, and the quality and the yield-increasing effect of the detoxified sweet potatoes are ensured by quickly and efficiently detecting the potyviruses.

Description

[0001] technical field [0002] The invention relates to the technical field of bioengineering, in particular to a serological detection kit and a detection method for potato Y virus on sweet potatoes. Background technique [0003] Sweet potato feather mottle virus ( Sweet potato feathery mottle virus , SPFMV), sweet potato latent virus ( Sweet potato latent virus , SPLV), sweet potato G virus ( Sweet potato virus G , SPVG) and sweet potato vein mosaic virus ( Sweet potato vein mosaic virus , SPVMV) are the main viruses that harm sweet potato, and all four viruses belong to the genus Potatovirus ( Potyvirus ), 4 kinds of viruses or mixed with other viruses often infect sweet potato, resulting in reduced yield, poor quality and degraded species of sweet potato, causing serious harm to sweet potato production. There is no particularly effective chemical control method for sweet potato virus disease at present. The most effective way to control virus disease is to culti...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/558
Inventor 张振臣乔奇张德胜秦艳红田雨婷王永江王爽
Owner HENAN ACAD OF AGRI SCI
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