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LAMP kit for detecting canine parvovirus

A canine parvovirus and kit technology, which is applied in the field of warm nucleic acid amplification technology kits to achieve the effects of easy observation, simple operation and high reliability

Active Publication Date: 2010-01-13
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But there is no application of this method in the detection of canine parvovirus

Method used

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  • LAMP kit for detecting canine parvovirus
  • LAMP kit for detecting canine parvovirus
  • LAMP kit for detecting canine parvovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] 1. Specific DNA sequence search

[0067] 从美国基因数据库——GenBank检索获得多株犬细小病毒基因组序列,登录号分别为EU697385、FJ869139-FJ86913922、FJ222822、FJ005265-FJ005200、EU659120、EU483515、EF666063、DQ354068、AY742955、DQ182617、DQ340405、AB120728、AB437433 , EU483516, through sequence homology analysis using BLAST software, a specific conserved target sequence was found for LAMP primer design. The specific conserved target sequence is:

[0068] TGC ATC ATT GAT GGT TGC ATT AGA TAG TAA TAA TACTAT GCC ATT TAC TCC AGC AGC TAT GAG ATC TGA GAC ATTGGG TTT TTA TCC ATG GAA ACC AAC CAT ACC AAC TCCATG GAG ATA TTA TTT TCA ATG GGA TAG AAC ATT AAT ACCATC TCA TAC TGG AAC TAG TGG CAC ACC AAC AAA TATATA CCA TGG TAC AGA TCC AGA TGA TGC TCA ATT TTA TACTAT CGA AAA TTC TGT GCC A

[0069] 2. Design of primers

[0070] Using Primer 4.0 (http: / / primerexlorer.jp; Eiken Chemical Co.Ltd, Japan), six primers were designed for the target sequence, including two inner primers (FIP and BIP) and two outer primers (F3 and B3) and two Str...

Embodiment 2

[0091] Adding loop primers (LF, LB) can increase the sensitivity of the LAMP reaction.

[0092] A comparative experiment was carried out, two groups of experiments, one group added loop primers, one group did not add loop primers, the results are shown in Figure 6.

[0093] Figure 6 is LAMP adding loop primer ( Figure 6-A ) and no loop primer ( Figure 6-B ) affects the reaction sensitivity comparison; among them, the plasmid copy numbers of lanes 1 to 7 are 2.0×10 6 , 2.0×10 5 , 2.0×10 4 , 2.0×10 3 , 2.0×10 2 , 2.0×10 1 , 2.0; 8 is the negative control; M is DL2000 plus, obviously the amplification reaction of adding loop primers is obviously stronger than that of the group without adding loop primers.

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Abstract

The invention relates to the technical field of biology, in particular to an LAMP kit for detecting a canine parvovirus. The kit comprises six LAMP primers, wherein the LAMP primers and LAMP reaction liquid form a detecting system together. The kit can rapidly and sensitively detect various current canine parvovirus strains, has simple operation, low cost and favorable specificity, is quite suitable for field detection of export quarantine, food sanitation and livestock farms and is easy for generalization and application in a large range, and a reaction result is easy to observe.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a loop-mediated isothermal nucleic acid amplification technique (LAMP) kit for detecting canine parvovirus. Background technique [0002] Canine parvovirus (Canine Parvovirus, CPV) is a virus discovered by Eugster in the United States in 1978 from the feces of dogs suffering from hemorrhagic enteritis. Canine parvovirus has become an important pathogenic virus of canine animals with the characteristics of high morbidity, strong infectivity and high mortality in dogs. Canine parvovirus has become one of the most serious infectious diseases in my country's dog industry. The disease not only affects the life and health of domestic pets, but also endangers the development of my country's dog industry, fur animal breeding industry and wild animal protection industry. [0003] At present, the traditional detection methods of canine parvovirus include serological detection methods and path...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 李刚张坤范晓娟
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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