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Method for detecting maize chlorotic mottle virus (MCMV) by using QCM (Quartz Crystal Microbalance) sensor and special gold piece

A technology of chlorotic mottled virus and gold flakes, applied in the biological field, can solve the problems of affecting experimental results, complicated ELISA operation, and long time consumption

Inactive Publication Date: 2014-05-14
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These traditional methods have certain defects in the detection of MMCMV. The ELISA operation is complicated and time-consuming, and the detection results are affected by experimental supplies and operating procedures; real-time fluorescent RT-PCR is prone to contamination and affects the experimental results.

Method used

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  • Method for detecting maize chlorotic mottle virus (MCMV) by using QCM (Quartz Crystal Microbalance) sensor and special gold piece
  • Method for detecting maize chlorotic mottle virus (MCMV) by using QCM (Quartz Crystal Microbalance) sensor and special gold piece
  • Method for detecting maize chlorotic mottle virus (MCMV) by using QCM (Quartz Crystal Microbalance) sensor and special gold piece

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Embodiment 1, the preparation of the QCM gold sheet that detects corn chlorotic mottle virus

[0086] 1. QCM gold flake pretreatment

[0087] Take out the QCM gold sheet (12mm×20mm, 50nm gold-plated on one side, BioNavis Company, article number QSX301-Au-13052), first rinse the surface with MilliQ water, and then put the gold sheet into the mixed solution (the mixed solution is 30% ammonia water , 30% hydrogen peroxide and MilliQ water in a volume ratio of 1:1:5) in a small beaker. First set the temperature of the water bath to 75°C and heat it up to the set temperature, then put the small beaker into the water bath (note that the beaker should not come into contact with the heating tube of the water bath to prevent the beaker from breaking), and process for about 10 minutes . Take out the beaker, use flat-tipped tweezers to pinch out the gold flakes, wash the residual liquid on the surface of the gold flakes with MilliQ water, and blow dry the surface with nitrogen; ...

Embodiment 2

[0114] Embodiment 2, utilize quartz crystal microbalance sensor to detect corn chlorotic mottle virus

[0115] 1. Control detection of MCMV

[0116] 1. Sensitivity, repeatability and specificity detection

[0117] Sensitivity, repeatability and specificity of MCMV detection using 11-MUA modified QCM gold sheet with immobilized MCMV monoclonal antibody:

[0118] 1) Sensitivity and repeatability of detecting MCMV

[0119] Dilute corn chlorotic mottle virus protein with PBS buffer to 5 concentrations, namely 1 μg / mL, 5 μg / mL, 10 μg / mL, 50 μg / mL, 100 μg / mL, and then perform QCM detection. The method is as follows:

[0120] Fix the 11-MUA modified QCM gold sheet of the fixed MCMV monoclonal antibody obtained in Example 1 on the test sample platform, turn on the peristaltic pump, set the flow rate to 50 μL / min; turn on the electronic unit, and perform data connection with the computer; turn on the Qsoft software, set the temperature at 22°C. Put the sample inlet tube into PBS; a...

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Abstract

The invention discloses a method for detecting a maize chlorotic mottle virus (MCMV) by using a QCM (Quartz Crystal Microbalance) sensor and a special gold piece. The invention provides a method for preparing a QCM gold piece for detecting viruses, wherein the method comprises the following steps: (1) modifying a QCM gold piece together with 11-mercaptoundecanoic acid (11-MUA) and 3-mercapto propionic acid (3-MPA), so as to obtain a modified gold piece; (2) fixing an antiviral antibody on the modified gold piece, thereby obtaining the QCM gold piece for detecting the viruses. According to the method, shown by experiments, the detection on MCMV is researched by using a QCM biosensor; through comparing and analyzing modified QCM gold pieces of two kinds, namely QCM gold pieces independently modified by 11-MUA and QCM gold pieces together modified by 3-MPA and 11-MUA, the QCM gold pieces together modified by 3-MPA and 11-MUA are higher in detection sensitivity and better in specificity and can be applied to actual detection methods.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting corn chlorotic mottle virus by using a QCM sensor and a special gold sheet. Background technique [0002] Mazie chlorotic mottle virus (MMCMV) is a positive single-stranded RNA with a diameter of 30nm icosahedron, which belongs to the genus Mazie chlorotic mottle virus and a member of the tomato bushy dwarf virus family. It seriously harms the growth of as many as 15-19 kinds of grasses such as wheat, barley, sorghum, and oats. Maize chlorotic mottle virus is widely distributed in Argentina, the United States and other European and American countries, and five countries in Africa. Due to its wide distribution, the risk of being introduced into my country is gradually increasing. If diseased plants are found in the field, they can be rapidly spread through insect mediators, seeds, etc., causing a large area of ​​corn to become diseased, and corn leaves will app...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531
CPCG01N5/02G01N33/536G01N33/56983
Inventor 黄新徐江敏
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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