PCR diagnostic kit for porcine infectious pleuropneumonia
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A diagnostic kit, pleuropneumonia technology, applied in the field of porcine infectious pleuropneumonia PCR diagnostic kit, to achieve the effect of simple detection method, accurate detection result and good use effect
Inactive Publication Date: 2010-06-09
GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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There is a lack of a simple, fast, sensitive and accurate detection kit for porcine infectious pleuropneumonia in clinical samples
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[0027] Embodiments of the present invention: porcine infectious pleuropneumonia PCR diagnostic kit, which comprises 400 μ L of proteinase K at a concentration of 20 mg / mL, 1000 μ L of lysate, the solution is Tris-HcL50mM, EDTA2mM, NaCl100mM; 1500 μL of TE buffer, TE buffer includes a mixed solution of Tris-HcL100mM and EDTA1mM, the pH value of the mixed solution is 8.0; 250 μL of PCR enzyme, PCR enzyme is Tiangen Biochemical Technology (Beijing) Co., Ltd. PCR MasterMix product, its specific composition includes 10mM Tris -HcL, 50mM KCL, 1.5mM MgCL 2 , 25mM dNTP Mixture and 0.05U Poymerase / uL; 170μL ultrapure water, 50μL Marker DL2000, 40μL equal volume mixed primers P1 and P2, the concentrations of primers P1 and P2 were both 20μM, and the sequences of primers P1 and P2 were respectively for P1:
[0028] 5'-ATGAGGATTTGTTTCTCGGTGGTG-3', P2: 5-TATTTCCGTCCGGTTTATTCAGGTC-3; 20 μL of ultrapure water was used as negative control; 20 μL of recombinant pMD18-T-apxIVA plasmid was used...
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Abstract
The invention discloses a PCR diagnostic kit for porcine infectious pleuropneumonia, which comprises 400mu L of proteinase K with the concentration of 20mg/mL, 1000mu L of cracking solution, 1500mu L of TE buffer solution, 250mu L of PCR enzyme, 170mu L of ultra-pure water, 50mu L of MarkerDL2000, 40mu L of primer P1 and primer P2 which are mixed in the same volume and have the same concentration of 20mu M, 20mu L of negative control and 20mu L of positive control. By optimizing the PCR reaction conditions, the invention develops a PCR kit for detecting the actinobacillus of the porcine infectious pleuropneumonia; and by comparing the PCR detection result with the negative control and the positive control, detection conclusions can be obtained, thereby achieving the aim of quickly detecting whether a sample has porcine infectious pleuropneumonia or not. The invention has accurate detection result, quick and sensitive detection process, simple detection mode and good using effect.
Description
technical field [0001] The invention relates to a kit, in particular to a PCR diagnostic kit for porcine infectious pleuropneumonia. Background technique [0002] Porcine Contagious Pleuropneumoniae (PCP) is a porcine respiratory disease caused by Actinobacillus Pleuropneumoniae (APP), characterized by hemorrhagic, necrotizing pneumonia and chronic fibrinous pleuropneumonia. Pigs die or grow poorly, causing huge economic losses to the swine industry around the world (Sebunya T N K et al., 1983, Macinnes J L et al., 1988, Gram T et al., 2000). It became popular in our country in the 1980s, and has become widely popular in recent years and has become an upward trend. According to whether it depends on NAD, it can be divided into two biotypes: biotype I and biotype II. The serotypes 1, 5, 9, and 10 of biological type I are the most pathogenic. According to statistics, the popular serotypes in my country are 1, 2, 3, 5, 7, 10 and other types, with 1, 3, and 7 being the main t...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/01
Inventor 蔡一鸣徐景峨谭诗文余波王璇任荣清
Owner GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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