基因2型牛病毒性腹泻病毒的分离、鉴定-兽医学;预防兽医学专业论文.docxVIP

Abstract1．RrI二PCR Abstract 1．RrI二PCR method髂tabljshed to detect aHd djstinguish BVDv-l and BVDv-2 B硒ed the GeIlBank data of me 5。岫仃anslated Ie垂。船(5’UTR)l【110wfl inⅡ1e GenBalllc fbm BVD、，fbr both BVDv．1 and BVDv．2 isoJates，n们o pa砥ofprimefs、他re pu斛Jy deSjgned to ampljfy spec讯cally 5‘UTR fh掣nelns f如m only BVDV．1，or oIlly BVDV之，and bom BVDV-l卸d BVDV-2， respcctively Based t11e above e解h pair of spec墒c pfinle塔fbr tlle r钾erse-缸锄scription polymemse chain 嘲cnon(R1：PCR)，ref确ce straills NADL(BⅦIV-1)柚d 890(BⅦIV-2)werc di脑e11tiated，The diseased tissue柚d orgaf塔(spIeen，mesentery lyHlph nodc，hean，cannon bom medu¨a，et．a1)f如m clinical c弱es of c眦Ie probabIy妯voIved in BvDv-2 infec6伽werc detected by d坤RT-PCR es诅bljshed above llsjngⅡ他specmc prime培for BVDV-2，the resuⅡs showed that 8 of 18(44％)samples we咒BVDv．2 rcla饽d infbcbon． 2．Isolation and identi傲a廿I阻of BVDV二2 T1le BVDV_2 positive sampIes by盯PCR∞m tlⅡ优地面oIls of XillJiall磊QinHai。sh孤D0ng懈 cho卵锄d inoculated into the MDBK ce¨s f研tlle、，ims isolaboIl，respec6vely Ancr 5～8 blind p豳sa孵iIl n碥cel{s．BVDV-2 mt毡cted MDBK were detected bv RT．．PCR．3 6eld isoIates of BVDV．2＆om d艟three ∞gio雎des嘶bcd above w嬲isolated，named XH)4 isolate，OH-92isolatc，SD-06 isolate respectively Und盯the light嘶croscope，jsolatesⅪ—04蛐d SDⅢ6 produce ce¨path02eIlic e腧t(CPE)includj雌 l啪y vacuolIlS，celIs gradtIaIIy losing i伍capability of aclIlerence，eventually bec砌ng mesh lcading to CPE．But iS01ate OH一92 h鹅∞visible CPE iJl MDBK ceU lines．hldirect i啪unonuorescencc嬲sav with (he specj矗c日uoresc曲ce reco辨ized in the cytopl鹪m showed 3 6e“isolatcs“BVDv-2把actcd with the 锄6．mou∞IgG of isolatc 890(BVDV之genotype)蛔_sed-E2 fIlsion肿teill．Under tIle electmn Inicroscope I啪y vims—like panicles wim envel叩e，round，a加彻d 60 in dian坨ter were obsenred in the uI自m-thin ∞ction of the endoDla釉11ic Teticulum f如mⅡ掂MDBK cells inf缸ted 3丘eJd isolates of BVDV．2．Same V确om、慨found by eIectron IIlicroscope using negatiVe s蜘ning Of phosphomngstic acid，which啪 purified f如m nle above infccted cells t11|rou曲出e procedure of tt