Real time-loop-mediated isothermal amplification (RT-LAMP) detection primers for performing differential diagnosis on canine distemper virus wild strains and vaccine strains and application of primers

A RT-LAMP, canine distemper virus technology, applied in DNA/RNA fragment, recombinant DNA technology, microbial determination/inspection, etc., can solve the problem that CDV wild virus strain and vaccine strain cannot be identified, and achieve high sensitivity , good repeatability and specificity

Inactive Publication Date: 2012-07-18
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
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Problems solved by technology

The RT-LAMP method for CDV has been reported, but none of them can be used for the identification of CDV wild strains and vaccine strains

Method used

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  • Real time-loop-mediated isothermal amplification (RT-LAMP) detection primers for performing differential diagnosis on canine distemper virus wild strains and vaccine strains and application of primers
  • Real time-loop-mediated isothermal amplification (RT-LAMP) detection primers for performing differential diagnosis on canine distemper virus wild strains and vaccine strains and application of primers
  • Real time-loop-mediated isothermal amplification (RT-LAMP) detection primers for performing differential diagnosis on canine distemper virus wild strains and vaccine strains and application of primers

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Experimental program
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Embodiment 1

[0029] 1 Materials and methods

[0030] 1.1 Virus strains and materials to be tested

[0031] Asia-I and Asia-II CDV field strains, CDV attenuated vaccine strains, CPV strains, canine adenovirus type I (CAV-I) strains, canine adenovirus type II (CAV-II) strains and rabies virus (RV ) strains were preserved by the Economic Animal Group of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences.

[0032] The materials to be inspected include 56 samples of clinical cases sent for inspection from different farms in Northeast China.

[0033] 1.2 Primer design and synthesis

[0034] Referring to the CDV H gene sequence in GenBank and using the online software Primer Explorer V4 as an aid, RT-LAMP primers were designed, including 2 outer primers F3 / B3 and 2 inner primers FIP / BIP (Table 1).

[0035] Table 1 RT-LAMP primer sequence

[0036]

[0037] 1.3 Viral RNA extraction and cDNA synthesis

[0038] 1.3.1 Extraction of RNA

[0039]Viral RNA was extract...

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Abstract

The invention discloses a set of real time-loop-mediated isothermal amplification (RT-LAMP) primers for differentiating canine distemper virus (CDV) wild strains from vaccine strains and application of the primers. The RT-LAMP primers comprise a pair of outer primers and a pair of inner primers, wherein the sequences of the outer primers are shown as SEQ ID NO:1 and SEQ ID NO:2 respectively; and the sequences of the inner primers are shown as SEQ ID NO:3 and SEQ ID NO:4 respectively. When the primers are used for detecting various genotypes of CDV wild strains according to an RT-LAMP detection method, the detection results are positive; and when the primers are used for detecting canine distemper vaccine strains, canine parvoviruses and other common canine viruses according to the RT-LAMPdetection method, the detection results are negative. Due to application of the primer sequence by the RT-LAMP detection method, the CDV wild strains and the CDV vaccine strains can be differentiatedaccurately; and the invention has the advantages of high specificity, high sensitivity, high repeatability and the like.

Description

technical field [0001] The invention designs a set of primers for detecting virus strains, in particular relates to a set of RT-LAMP primers for detecting canine distemper virus wild strains, and belongs to the detection field of canine distemper virus. Background technique [0002] Canine distemper virus (CDV) belongs to Paramyxoviridae, Paramyxovirinae, Morbillivirus. CDV is an enveloped single-strand, negative-strand, non-segmented RNA virus with a genome length of 15 690 bp, mainly encoding nucleocapsid protein (N), phosphoprotein (P), matrix membrane protein (M), fusion Protein (F), hemagglutinin (H) and large protein (L) six structural proteins. At present, it is recognized that CDV has only one serotype, but according to the principle that strains with H gene amino acid sequence homology higher than 95% can be classified as the same genotype, CDV can be divided into Asia-I and Asia-II (Asia-II), European type (Europe), American type (USA or America), Arctic type (Ar...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 刘大飞张洪英曲连东戚亭刘春国
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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