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Method for detecting canine influenza

A detection method, a detection dog technology, applied in the direction of biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of many influencing factors, high price, ELISA false positive, etc., achieve low quality requirements, improve specificity, highly specific effect

Inactive Publication Date: 2010-09-22
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although ELISA detection kits are also used in clinical practice, ELISA detection has certain false positives, and its price is very expensive
Another disadvantage of ELISA detection is that the detection kits are all batch detection, which is not very beneficial to the sporadic outbreak of the H3N2 subtype canine influenza disease, which will increase the considerable cost on the basis of its expensive price
As a result, the cost of testing dogs is too high, and it is difficult for animal hospitals and pet owners to accept
Furthermore, when a single dog or a few dogs are tested by ELISA, it will cause a serious waste of kits
In addition, there are many and complex factors affecting the ELISA detection operation, which also cause a certain degree of unreliability in the results.

Method used

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  • Method for detecting canine influenza
  • Method for detecting canine influenza
  • Method for detecting canine influenza

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Take a nasopharyngeal swab from a clinical case dog, put it into a centrifuge tube filled with 750 μL of normal saline, squeeze it several times, and use the supernatant as the virus solution for RNA extraction.

[0028] The specific operation steps are as follows:

[0029] Add 750 μL Trizol Ls Reagent to 250 μL virus solution, shake well at room temperature (15-30°C) for 5 minutes, then add 200 μL chloroform, shake well at room temperature (15-30°C) for 1-2 minutes, centrifuge at 12000 g, 4°C for 10 minutes , absorb about 500 μL of supernatant, add 500 μL of isopropanol, centrifuge at 12,000 g, 4°C for 10 min, discard the supernatant, add 1,000 μL of 70% DEPC ethanol, centrifuge at 7,500 g, 4°C for 5 min, and place in an ultra-clean bench After fully drying, add 20 μL DEPC water and store at -20°C.

[0030]Add 1 μL of random primers to 5 μL of the virus RNA extracted above, and then react in a 70° C. water bath for 5 minutes, and then ice-bath for 10 minutes. Then ad...

Embodiment 2

[0035] As in Example 1, but the samples were nasopharyngeal swabs from healthy dogs.

[0036] The result is as figure 2 As shown: the positive sample has the target band; the negative control has no target band; the test sample has no target band, and it is judged as negative.

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Abstract

The invention belongs to the field of biotechnology and particularly relates to a method for detecting canine influenza. In the invention, a PCR detection method is adopted, and a PCR reaction system comprises the following materials: primers having sequences SEQ ID No.1 and SEQ ID No.2 respectively, as well as dNTPs, Ex Taq, 10*Ex Taq buffer and cDNA templates. At the end of a PCR reaction, whether a target strip exists is detected by electrophoresis and ultra-violet analysis, so as to determine if a dog catches canine influenza. The detection method provided by the invention has high sensitivity and specificity, is simple, convenient and quick in operation, has low requirements on the quality of initial materials and is convenient to use in clinic.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for detecting canine influenza. Background technique [0002] Canine influenza is caused by various influenza A viruses, which were discovered in 2004 to cause canine influenza. Dogs do not have natural immunity to this type of virus. Therefore, the infectious disease may spread rapidly among dogs. It is a disease with a high incidence, mainly manifested as eye and nose secretions, sneezing, coughing and other respiratory symptoms. , It is also easy to be confused with respiratory diseases such as canine distemper and parainfluenza in clinical manifestations. H3N2 subtype canine influenza virus has been detected and successfully isolated in dogs in my country. However, at present, there are no effective vaccines and drugs for the prevention and control of canine influenza at home and abroad. The main means of controlling the disease is through early detection o...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 李守军师志海焦培荣远立国
Owner SOUTH CHINA AGRI UNIV
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