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Primer, amplification reaction liquid, kit and detection method for LAMP detection of canine parainfluenza

A technology of canine parainfluenza and amplification reaction, which is applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/testing, etc., can solve the problems of easy false positive results, low sensitivity, expensive equipment, etc. Achieve the effect of broad market application prospect, high scalability and low cost

Inactive Publication Date: 2021-05-25
爱若维生物科技(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection methods for antibodies include immunofluorescence method, ELISA, neutralization experiment and hemagglutination inhibition test, etc., but these methods have great limitations, the equipment required for RT-PCR is expensive, the operation is cumbersome, and the sensitivity of colloidal gold test strips is low. Weak specificity, prone to false positive results, ELISA is used to detect antibodies, the detection time is long and the sensitivity is not strong

Method used

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  • Primer, amplification reaction liquid, kit and detection method for LAMP detection of canine parainfluenza
  • Primer, amplification reaction liquid, kit and detection method for LAMP detection of canine parainfluenza
  • Primer, amplification reaction liquid, kit and detection method for LAMP detection of canine parainfluenza

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042]A primer for the detection of canine parainfluenza LAMP, including two outer primers F3-1 and B3-1, two inner primers FIP-1 and BIP-1, and a loop primer LB-1; the sequence of the primer is as follows As shown in Table 1,

[0043] Table 1 The sequence list of the first set of primers for LAMP

[0044]

Embodiment 2

[0046] A primer for the detection of canine parainfluenza LAMP, including two outer primers F3-2 and B3-2, two inner primers FIP-2 and BIP-2, and a loop primer LB-2; the sequence of the primer is as follows As shown in Table 2,

[0047] Table 2 LAMP second primer sequence list

[0048]

Embodiment 3

[0050] A primer for the detection of canine parainfluenza LAMP, including two outer primers F3-3 and B3-3, two inner primers FIP-3 and BIP-3, one loop primer LB-3 and one loop primer LF; The sequences of the primers are shown in Table 3,

[0051] Table 3 LAMP third primer sequence list

[0052]

[0053] The present application also provides an amplification reaction solution for the detection of canine parainfluenza LAMP, the reaction solution includes but not limited to the primers for the detection of canine parainfluenza LAMP described in Example 1-Example 3. As a preferred embodiment of the present application, the reaction solution also includes Thermopol buffer, dNTP, MgSO 4 , betaine and Bst DNA polymerase, wherein: the Thermopol buffer contains Tris-HCl, KCl, (NH 4 ) 2 SO 4 , MgSO 4 and Triton X-100.

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Abstract

The invention relates to a primer, an amplification reaction solution, a kit and a detection method for LAMP (loop-mediated isothermal amplification) detection of canine parainfluenza virus, the kit and the detection method can accurately distinguish samples containing canine parainfluenza virus and samples not containing canine parainfluenza virus, namely after LAMP specific detection, the specificity of the designed primer is good. According to the kit and the detection method, the canine parainfluenza virus can be quickly and accurately detected, results are observed through visual inspection or instruments, data processing is simple, cost is low, and the kit and the detection method have important significance on clinical detection, are particularly suitable for livestock farms, are economical in detection and can be widely popularized.

Description

technical field [0001] The invention relates to the technical field of animal inspection and quarantine, in particular to a primer, amplification reaction solution, kit and detection method for canine parainfluenza LAMP detection. Background technique [0002] Canine parainfluenza (CPIV) is caused by parainfluenza virus type Ⅱ and is a viral infectious disease in dogs with acute respiratory inflammation as the main symptom. Parainfluenza virus type Ⅱ is a member of the family Paramyxoviridae and the genus Paramyxovirus. It can also cause acute encephalomyelitis and hydrocephalus, with clinical manifestations such as hindquarter paralysis and ataxia. The surface of its virus particles contains hemagglutinin and neuraminidase, which can agglutinate red blood cells of human type O, chicken, guinea pig, rat, rabbit, dog, cat and sheep. The resistance of the virus to physical and chemical factors is not strong. In a protein-free liquid medium, it almost loses its infectivity at ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844
Inventor 王蓉蓉冯小龙孙云雷
Owner 爱若维生物科技(苏州)有限公司
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