Loop-mediated isothermal amplification (LAMP) diagnostic kit for porcine circovirus type 2

A porcine circovirus and diagnostic kit technology, applied in the field of kits, can solve the problems of low sensitivity, time-consuming and laborious, complicated operations, etc.

Active Publication Date: 2014-05-28
GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, conventional pathogenic isolation and serological detection methods have the disadvantages of complex operations, time-consuming and labor-intensive, low sensitivity, etc. PCR methods require expensive PCR instruments

Method used

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  • Loop-mediated isothermal amplification (LAMP) diagnostic kit for porcine circovirus type 2
  • Loop-mediated isothermal amplification (LAMP) diagnostic kit for porcine circovirus type 2
  • Loop-mediated isothermal amplification (LAMP) diagnostic kit for porcine circovirus type 2

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Embodiment Construction

[0058] Embodiment of the present invention: porcine circovirus type 2 LAMP diagnostic kit, which includes 250 μL of LAMPmix buffer, 20 μL of 0.5 U / μL Bst DNA polymerase, 150 μL ultrapure water, the volume of inner primer 1 and inner primer 2 is 40 μL, the concentration is 40 pmol / L, the volume of outer primer 1 and outer primer 2 is 40 μL, the concentration is 5 pmol / L, 20 μL Negative control and 20 μL positive control; the sequence of outer primer 1 is: 5 ， -TGGAGCTCCTCGATCTCAAG-3 ， ; The sequence of the outer primer 2 is: 5 ， -GCCCCACAATGACGTGTAC-3 ， ; The sequence of inner primer 1 is: 5 ， -GCAACGGTCACCAGACTCCCGACAACGGAGTGACCTGTC-3 ， ; The sequence of inner primer 2 is: 5 ， -AGAGCAGCACCCTGTAACGTTTACGCTTCTGCATTTTCCCG-3 ， ; Negative control is ultrapure water; Positive control is porcine circovirus type 2 strain DNA; LAMP mix buffer includes 3 mmol / ul MgCl 2 , 500pmol / ul dNTP 2.0μL and 25mmol / μL Tris-HCl, the pH value is 8.3.

[0059] SEQUENCE LISTING

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Abstract

The invention discloses a loop-mediated isothermal amplification (LAMP) diagnostic kit for porcine circovirus type 2. The diagnostic kit comprises 250muL of LAMPmix buffer solution, 20muL of (i)Bst(/i) deoxyribonucleic acid (DNA) polymerase, 150muL of ultrapure water, 40muL of mixed inner primer 1 and inner primer 2, of which the concentration is 40pmol/L, 40muL of mixed outer primer 1 and outer primer 2, of which the concentration is 5pmol/L, 20muL of negative control and 20muL of positive control. Two pairs of primers are designed and synthetized according to a porcine circovirus type 2(PCV-2) ORF2 gene sequence in GenBank, an LAMP diagnostic method for the porcine circovirus type 2 is built by optimization of the LAMP condition, and the sensitivity and specificity tests, and a simple and fast method is provided for rapid diagnosis of the porcine circovirus type 2 on the spot. Thus, quick quarantine needs of the basic level on the spot are met.

Description

technical field [0001] The invention relates to a kit, in particular to a PCR diagnostic kit for giant salamander pathogenic Aeromonas hydrophila. Background technique [0002] Porcine circovirus type 2 is the smallest animal virus found so far and can cause multisystemic wasting syndrome in weaned piglets, reproductive failure in pregnant sows, respiratory disease in weaned pigs and finishing pigs, porcine dermatitis and nephritic syndrome, congenital Sexual tremor and other diseases have caused great economic losses to pig production in my country. [0003] At present, the main detection methods include virus isolation and identification, agar diffusion test, trace serum neutralization test, ELISA, and PCR methods. Moreover, the conventional pathogenic isolation and serological detection methods have the disadvantages of complicated operation, time-consuming and laborious, and low sensitivity. The PCR method requires an expensive PCR machine. Loop-mediated isothermal ampl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6844C12Q1/70C12Q2531/119
Inventor 余波冉懋韬谭诗文
Owner GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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