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Tobacco potato Y virus field rapid detection test paper preparation method

A technology for detecting test strips and potatoes, which can be applied to measurement devices, instruments, scientific instruments and other directions, and can solve the problems of low titer of antiserum and unsatisfactory detection effect.

Inactive Publication Date: 2008-07-02
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, both of these methods require the extraction and purification of viruses or nucleic acids in the samples, and equipment such as ultra-high-speed centrifuges and electron microscopes are required. Although the specificity of serological methods is relatively strong, when the virus concentration is low, the antiserum prepared is less effective. The price is not high, the detection effect is not ideal

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Antibody preparation: Preparation of rabbit polyclonal antibody against Tobacco Potato Y virus: Dilute the antigen (purified PVY) to 2mg / ml with sterile saline, and then add Freund's adjuvant. The first time is complete adjuvant, and then incomplete adjuvant is used. Agent. The amount of antigen injection is 0.1~0.2mg / kg, 2~3Kg rabbits are selected, and the intravenous injection is done in three times. The amount of antigen injection is 0.03~0.1mg each time, and the interval is 10d. Bloodletting was performed 10 days after the last injection. Animals were fasted for 12 hours before bleeding, and 40-60ml of blood was used for ear vein bleeding. After collection, the blood cells were removed by centrifugation, and stored at minus 20°C after purification. Goat anti-rabbit antibodies are also prepared.

[0014] Preparation of colloidal gold particles: Take 0.6 ml of 1% chloroauric acid in 30 ml of ultrapure water and boil it. Quickly add 0.9ml of 1% sodium citrate pre-warmed at...

Embodiment 2

[0019] Example 2: In this example, when preparing colloidal gold particles: 0.6 ml of 1% chloroauric acid was heated and boiled in 30 ml of ultrapure water. Quickly add 0.9ml of 1% sodium citrate pre-warmed at 37°C at a time, the solution gradually changes from blue to purple-red, boil for 3min, and add water to 30ml. After cooling to room temperature, the pH was adjusted to 7.5 with 0.2mol / L potassium carbonate. Others are the same as in Example 1.

Embodiment 3

[0020] Example 3: In this example, when preparing colloidal gold particles: 0.6 ml of 1% chloroauric acid was heated and boiled in 30 ml of ultrapure water. Quickly add 0.9ml of 1% sodium citrate pre-warmed at 37°C at a time, the solution gradually changes from blue to purple-red, boil for 4min, and add water to 30ml. After cooling to room temperature, the pH was adjusted to 7.4 with 0.2mol / L potassium carbonate. Others are the same as in Example 1.

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PUM

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Abstract

The invention discloses a preparation method of test strip for rapid field detection of tobacco potato virus Y, which comprises the following steps of: preparing an antibody, preparing colloidal gold, preparing a colloidal gold-antibody conjugate, preparing a nitrocellulose filter, and preparing a colloidal gold chromatographic test strip. The prepared test strip for rapid field detection of tobacco potato virus Y according to the technical proposal has an optimum detection temperature of 15-35 DEG C, sample detection amount of 0.1-0.3g and tobacco sample area of 3-5cm<2>. The detection method comprises charging tobacco leaf in a plastic bag containing buffer, grinding, immersing the test strip in the buffer, reacting for 30min, and observing the result. The false positive rate is less than 1%. The invention has the advantages of convenient application and high reliability; and is especially suitable for rapid field detection.

Description

Technical field [0001] The invention belongs to detection materials, and particularly relates to a method for preparing a test strip for rapid field detection of tobacco potato Y virus. Background technique [0002] Tobacco potato virus Y disease, also known as tobacco vein band disease, is distributed in all tobacco areas in the world. In recent years, my country’s Huanghuai, Southwest, Northeast and other smoking areas have tended to aggravate the occurrence, and have caused relatively large economic losses in some smoking areas. The nucleic acid of Potato virus Y (PVY) is a positive-strand ssRNA with a length of about 10 kb. It mainly overwinters on farmland weeds and Solanaceae plants. In warm areas, it continuously reproduces and infects on the host; through aphids and sap Toxins are transmitted through multiple ways such as friction and grafting. Under natural conditions, aphids are still the main source of transmission; clear veins appear on the new leaves at the early sta...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/569
Inventor 高玉千邱立友梁振普戚元成
Owner HENAN AGRICULTURAL UNIVERSITY
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