Method of synchronous distinguishing newcastle disease virus and vaccine virus and identifying virulence and genotype
A newcastle disease virus and vaccine virus technology, applied in the field of automated analysis software, can solve the problems of small number of verification samples, insufficient theoretical foundation, and difficult application
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Embodiment 1
[0116] Embodiment 1-isolate the detection of Newcastle disease strain
[0117] 1. Virus isolation and identification.
[0118] According to the method of "Diagnostic Test and Vaccine Standard Manual" (OIE, 1996), 51 strains of Newcastle disease virus were isolated from geese, pigeons, chickens and ostriches.
[0119] 2. Extraction of viral nucleic acid RNA
[0120] (1) Take 100 μl of allantoic fluid to be tested.
[0121] (2) Add 900 μl TRIzol reagent (Gibco-BRL company), mix well, and keep at room temperature for 5 minutes;
[0122] (3) Add 0.2ml or appropriate amount of chloroform, vortex and mix well, then let stand at room temperature for 3 minutes;
[0123] (4) 12000g, 4°C, centrifuge for 15 minutes;
[0124] (5) Carefully draw the supernatant into a new 1.5ml centrifuge tube;
[0125] (6) Add 0.5ml or an appropriate amount of isopropanol, vortex and mix well, then place at room temperature for 10 minutes;
[0126] (7) 12000g, 4°C, centrifuge for 10 minutes;
[012...
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