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SERS-based newcastle disease virus detection kit and detection method thereof

A technology of Newcastle disease virus, detection kit, applied in the field of spectroscopy and molecular diagnosis

Active Publication Date: 2015-11-25
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is a lack of research on the application of Raman spectroscopy to the detection of Newcastle disease virus with high-efficiency kits.

Method used

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  • SERS-based newcastle disease virus detection kit and detection method thereof
  • SERS-based newcastle disease virus detection kit and detection method thereof

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Embodiment 1

[0026] A kind of Newcastle disease virus detection kit and detection method thereof based on SERS (Surface Enhanced Raman Spectroscopy Technology) are provided in the present invention, and concrete steps are as follows:

[0027] (1) Preparation of nano-gold: 1mL of 1% chloroauric acid (HAuCL 4 ) was dissolved in 100mL ultrapure water, and then 2mL of mass fraction 1% sodium citrate solution (Na 3 C 6 h 5 0 7 ), about 1 mL of 0.1M potassium carbonate (K 2 CO 3 ) to adjust the pH to 7; the above solution was heated at 50° C.-100° C. for 20-30 minutes in a condensing reflux device until the color turned dark purple. The size of the obtained nanoparticles measured by photon correlation spectroscopy is about 20nm, and the concentration is 9×10 11 Particles / mL;

[0028] (2) Preparation of 3,3'-dithiobis(6-nitrobenzoic acid)imidosuccinate: take a 100mL round-bottomed two-necked flask, add 50mL of dry tetrahydrofuran under nitrogen, and add 0.5g of 5 , 5'-dithiobis(2-nitroben...

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Abstract

The invention discloses an SERS-based newcastle disease virus detection kit and a detection method thereof, and belongs to the field of spectroscopy and molecular diagnosis. The SERS-based newcastle disease virus detection kit comprises a raman probe solution and a nitrocellulose film; the raman probe solution is a solution obtained by coupling nano gold, 3,3'-dithio di(6-nitrobenzoic acid) di-succinic acid imido ester complex and a newcastle disease antibody. The detection method comprises the following steps: dropping a newcastle disease virus on the nitrocellulose film, incubating the nitrocellulose film and the raman probe coupled with the antibody after closing and washing to form an immune complex with a 'solid phase antigen-labeled antibody' half sandwich structure on the nitrocellulose film, and finally performing raman spectrum detection on the immune complex so as to identify an enhanced raman molecular signal of a material to be identified with high specificity. The detection kit and the detection method which are disclosed by the invention are high in specificity, and the detection limit of the newcastle disease virus is 10<3>PFU / mL, and the sensitivity is high.

Description

technical field [0001] The invention belongs to the field of spectroscopy and molecular diagnosis, and in particular relates to a detection kit for Newcastle disease virus based on SERS (surface-enhanced Raman spectroscopy) and a detection method for others. Background technique [0002] Newcastle disease (Newcastledisease, ND), also known as Asian chicken plague, is a highly contagious, acute septic avian infectious disease caused by Newcastle disease virus (NDV) with respiratory tract and digestive tract mucosal bleeding as typical lesions. Higher morbidity and mortality are listed as legally notifiable animal diseases by the World Organization for Animal Health (OIE). ND has widely existed in all kinds of poultry flocks for many years, causing huge economic losses to the breeding industry in our country. At present, the existing detection techniques for Newcastle disease virus mainly include: virus isolation and culture, hemagglutination inhibition (HI) test, enzyme-link...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65G01N33/569
Inventor 任涛谭阳通冯敏莎
Owner SOUTH CHINA AGRI UNIV
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