Study On Direct Immunofluorescent Method To Detect GPMV-Ⅰ And Distribution Of The Viral Antigen In Artificially Infected Gooses With GPMV-Ⅰ By Means Of DIF

The anti-GPMV-â… IgG was obtained in healthy gooses after being inoculatedwith GPM virus allantoic fluid which was Inactivated with Formalin, and then waslabelled with fluorescein isothiocyanate(FIYC). Driect ImmnohistochemicalMethod was successfully developed and standardized to detect the GPMV-â… antigen in artificially infected gooses. It proved that the DirectImmunofluorescence Staining Method (DISM) has simple opreation, highsensitivity and specificity by absorption assay, replacement assay and positive andnegative c6ntrol assay. Through optimization, DISM could get excellent resultsunder the follow conditions: Frozen sections fixed with Acetone, keeping slices in4~C 30min, treating clean slices by PLL (Poly-L-Lysine), the dilution ratio of FITClabelling goose anti-PGMV-1 is 1:16, repairing antigen by Trypsin for 20min,and staining slices with FITC-IgG for 30min.DISM is a specific, sensitive and visual staining method. It can be applied todiagnose the GPMV-â… , to detect the distribution of GPMV-â… antigen in tissuesof the infected gooses, and in the retrospective diagnosis in formalin-fixed tissues.Only special positive signals emerged in the tissues of Goslings infected byGPMV-â… , but not emerged in the liver tissues infected by GPV, goose E. coli,Salmonella, Pasteurella. We systematically investigated the consecutivedevelopmental distribution regularity of goose Paramyxovirus in artificallyinfected goslings respectively. The result of inoculated group in diferent hourspost inoculation: GPMV-â… antigen can be detected from kidney and liver at 8hpi; GPMV-â… antigen can be detected from liver, spleen, lung, kidney, duodenum,Pancreas, thymus at 12hpi; GPMV-â… antigen can be detected from nine tissuessuch as liver, lung, proventriculus, and so on at 24hpi; GPMV-â… antigen can bedetected from all tissues excepted for muscle, myocardium and cerebrum from 96hpi; GPMV-â… antigen can not be detected from gullet and rectum at 192hpi, andthe positive extent decreased gradually in other tissues; GPMV-â… antigen still canbe detected from liver and kidney at 288hpi. The subsequence of positive rate ininoculated group: liver(17/22), kidney(16/22), trachea(14/22), lung(14/22),duodenum(13/22), bursa of Fabricius(12/22), Pancreas(12/22), spleen(11/22),thymus(11/22), proventriculus(10/22), gullet(6/22), rectum(5/22). The virusantigen can't be detected from any tissues of goslings at 4hpi and 20dpi by theroute of inoculation. The distribution rate of inoculation at 96hpi was fastest. Themax detection rate was the liver among the detected tissuses. During this research,we didn't detected the GPMV-â… antigen from muscle, myocardium and cerebrumusing DIF method all along.