(P seudo rab ies, PR ) (A u jeszky, s D isease, AD ), (Pesuderab ies viru s, PRV )

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1 34 9 ( ) V o l. 34 N o Jour. of N o rthw est Sci2T ech U niv. of A gri. and Fo r. (N aṫ Sci. Ed. ) Sep W G Ξ 1, 1, 1, 1, 1, 2 (1, ; 2, ) [ ]ge2el ISA (P seu2 do rabies virus, PRV ),, PRV %, %, 0; PK215,,, CPE 20 h ;, PCR PRV g E 612 bp PRV, PRV W G []; ; [ ] S [ ] A [ ] (2006) (P seudo rab ies, PR ) (A u jeszky, s D isease, AD ), (Pesuderab ies viru s, PRV ) [1 ],,,, PRV, ;,,, 80% 100%,,,, 100%, 1, 1960,,,, PR,, [2 ], 24 [3 ],, [4 ],,, [5 ] PRV, PRV, PR, ge2el ISA,, PCR, PRV PR PK215 ; PRV Bartha2k61 (g E ) PRV M in2a, ; , , BAORAD ;, Eppendo rf ; ge2el ISA ( : 22g03g05), Ξ [ ] [ ] (2004K022G3201) ; ( ) [ ] (1972- ),,,,, [ ] (1963- ),,,,,

2 32 ( ) 34 ID EXX, 96,, (HR PO ), TM B,,, ; 2 PCR T aqm ix 100 bp DNA L adderm arker ; K (SD S) ( ) ; DM EM G IBCO, GenBank PRV (: BK001744), 1, P1: 5 2AA T A T G CGG CCC T T T CT G CT G23 ; P2: 5 2AA T CA C AAA GAA CA C GGC CC23, 612 bp, bp, , 6, ge2el ISA ge 1. 2 PR ge2el ISA 187, SgN ( OD 650 g OD 650 ) : SgN 0. 6, ; Sg N > 0. 7, ; 0. 6< SgN 0. 7, 1. 3 PRV W G ,, IU gml ΛggmL H ankπs 5, 4 4 h, , rgm in 30 m in,, Λm,, % PK215,, 2 ml, 37 1 h, m in,,, 2% 100 IU gml 100 ΛggmL DM EM 8 ml ; 1 8 ml DM EM, 5% CO % (CPE) 5, CPE, , 2 2 ml g ; 2 2 ml g ; 2 H ankπs 2 ml g,, PCR (1) DNA [6 ],,, PRV Bartha2K61 PRV M in2a 3, rgm in 30 m in, rgm in 20 m in, 400 ΛL T E, K 100 Λgg ml, SD S 10 ggl, 55 1 h, rgm in 5 m in,, T ris2 1, V ( ) V ( ) V () = , V ( ) V () = , DNA, 4 75% 2, 20 ΛL, PCR, - 20 (2) PCR [ 7 ] PCR PCR : 10 Λmo lgl P1, P2 1 ΛL, DNA 5 ΛL, T aq M ix ΛL, 5. 5 ΛL PCR : 96 5 m in; 96 1 m in 10 s, 65 1 m in, 72 1 m in, 35 ; m in; 4, PCR 10 ggl (3) PCR PCR, pm D 182T V icto r 5 ΛL DH 5Α, LB, h 4. 5 ml LB, h, H indg Kp ng PCR 10 ggl, DH 5Α , PRV %, %,

3 9 : W G 33, ; 1. 15% PRV, P ig farm 1 ge2el ISA, T able 1 ge2el ISA exam ination result of serum samp le from different p ig farm s N um ber of detection Po sitive num ber g% Po sitive rate Doubtful num ber A p ig farm in huxian county A p ig farm in yangling city A p ig farm in yulin city A p ig farm in xingp ing city A p ig farm in bao ji city g% Doubtful rate A p ig farm in heyang county To tal V alue of average CPE PK215, 5, 1 CPE, 48 h,,,,,,,, ( 1 A B ),, ( 1 C), CPE PK215 10, h, 80% CPE 20 h 1 PRV PK215 (100 ) A B.,,, ;C. PK215 F ig. 1 Cell cytopath ic effect (CPE) results of PK215 by PRV (100 ) A B. Infected PK215 cell line shrunk, aggregated, and detached from the culture system by PRV and the typ ical patho logy w ere observed; C. T he contro l PK215 cell h,, ; 48 h,, ; h, h, 2. 4 PCR PCR 2, PRV M in2a, 612 bp, ; PRV Bartha2K61 2 PCR M. DNA m arker; 1. ; 2. ; 3. M in2a ; 4. ; 5. Bartha2K61 ; 6. ddh 2O F ig. 2 A gro se gel electropho resis of PCR p roducts of PRV M. DNA m arker; 1. Cell culture; 2. Po sitive m aterial; 3. M in2a strain; 4. Cell contro l; 5. Bartha261 strain; 6. ddh 2O contro l

4 34 ( ) , H indg Kp ng 612 bp 2. 7 kb, PCR 612 bp, pm D 182T vecto r 3 PCR M. DNA ; 1. pm D 182T ; 2. ; 3. ; 4. PCR ; 5. PCR F ig. 3 Identification of the recom bined p lasm ids by PCR and restriction digestion M. 100 bp L adder DNA M arker; 1. pm D 182T V ecto r p lasm ids; 2. the recom bined p lasm ids; 3. restriction identification of the recom bined p lasm ids; 4. PCR of the recom bined p lasm ids; 5. PCR p roducts of PRV, PRV, PRV W G PRV, 5, 2000 PR %, % [5 ],,, PRV, PR %, 50% 40% PRV, PRV,,,, PR, 3. 2 PK215 PCR, PRV, PRV W G PRV W G PK215, h, h,,, [8 ], PRV W G,,, PRV 3. 3 PRV PCR 2,,, PCR ge ge, PCR [9 ] g24gEEC,, PRV 12 g E 2 g E 2EL ISA ; g E, g E 2EL ISA, g E 2EL ISA PCR, g E PCR, PRV PR g E, GeneBank 4 PRV g E 1, PRV g E PCR, g E Bartha2K61, [7 ] g E,, g E 2 EL ISA [10 ],, PRV,

5 9 : W G 35,,,, PCR, g E 2 EL ISA PCR,, PCR,, [ 1 ],. [M ]. 2. :, [ 2 ],. [M ]. :, [ ] [ 3 ]. [D ]. :, 2005: [ 4 ]. [J ]., 2002, 34 (10) : [ 5 ],,,. [J ]., 2001, 31 (4) : [ 6 ],,,. A [J ]., 1998, 29 (2) : [ 7 ],,,. PCR [J ]., 2004, 19 (6) : [ 8 ],. L Y [J ]., 2003, 23 (3) : [ 9 ] L aszlo zsak, Federico zuckerm ann, N ancy Sugg, et al. Gulycop ro tein g I of P seudo rabies virus p romo tes cell fusion and virus sp read via di2 rect cell to cell transm ission[j ]. J V iro l, 1992, 6: [ 10 ] D avid K, Sabrina L S, L ie2l ing W, et al. Evaluation of sero logical tests fo r the detection of p seudo rabies ge antibodies during early infec2 tion[j ]. V et M icro, 1997, 55: Iso lation of p seudo rab ies viru s Shaanx iw G strain and ep idem io logy investigation J IANG Yan-fen 1,YANG Zeng-q i 1, ZHU W e i-guo 1, W ANG Xu-rong 1, TIAN X iao-yan 1,W ANG Qing-a i 2 (1 Colleg e of A nim al S cience and T echnology,n orthw estern A & F U niversity, Y ang ling, S haanx i , Ch ina; 2 Z henba A nim al H usband ry and V eterinary S tation, Z henb a, S haanx i , Ch ina) Abstract: A bou t 187 serum samp les from 6 p ig farm s in som e region s of Shaanx i p rovince w ere detect2 ed w ith the g E 2EL ISA K it w h ich can distingu ish the vaccination from w ild2type viru s infection, and po rcine p seudo rab ies viru s w as iso lated from the purtenance and b rain of a 22day2o ld p ig and then the viru s w as i2 den tified m icrob io logically. T he resu lts show ed that the average po sitive rate w as %, the h ighest g E 2 an tibody of w ild2type PRV w as % and the low est w as 0; It infected PK215 cell line and cau sed it to sh rink, aggregate, and detach from the cu ltu re system in 20 h app rox im ately, and the inocu lated rabb its show ed classical clin ical sign s w hen challenged w ith the cu ltu re and po sitive m aterial. T hen the specific 612 bp bands by PCR of the iso lated viru s w ere ob tained. T hese data indicate that the iso lated viru s w as PRV and referred to as W G strain in th is repo rṫ Key words: p seudo rab ies viru s; iso lation and iden tification; ep idem io logy