利来w666备用Ʒ | PRODUCT
ҳ > >

һ к  ߷

2016-01-06 ҵѧ

1

1.1 硢Լʵ鶯Դ

ΪʵƵԸк¶ 磬 Ʒ ΪȾθ- Ըк硣RNA ȡԼйGenemark ˾Ըк IgA IgG ELISA Ϊʵ ƣϸԼ gibco hyclone ˾ ʵԴ ӱijģ

1.2  

ó ֣ʳ״кݡк۲첡仯ȡıá

1.3 ʵ Ҳԭѧ

СƷĥ3 ΣTRIzol ȡRNAʹͬʱPEDV M TGEVȾ θײN GARA Ⱥ״VP7 ĶRT-PCR Ʒ

1.4

ҪԸóѡĸ3  ͬ߳ ķЧʹǽ3 Ϊ飬飬顣 ߳1

 1

 صҪ ȫۣRT-PCR ԭѧĸPEDV Ŷ ĸ ʳк ȣЧ ʵۣͨELISA ⿹PEDV IgA 塢IgG 弰ͨ ккˮƽٴۣͳĸ7 d Ѿ ʡ

1.5 ELISA

ɼ3 ĸ 42 ݣ4 棬12 000 r/min 5 min ȥϲ ֬㣬ǹͷС ȡм 壬ʹʵ  Լ кIgAIgG 壬 Լ˵顣 ж S/P ֵΪIgA ˮƽı׼S/P ֵ≥ 0.148 ΪԣS/P ֵ 0.148ΪԡOD ֵΪIgG ˮƽı׼OD ֵ≥ 0.433 ΪԣOD ֵ 0.433 Ϊԡ

1.6 к

TCID50 PEDV-YN144  Һϡ 200 TCID50/50μLIJҺ96 ΢н 2 ϡͣ50 µL ϡ õIJҺȺ37 5% CO2 60 minͬʱ  գѪգ պϸ գ 2002020.2 TCID50 4 ݶȶա  ɺȥҺDMEM ϴ2 Σÿ׼200 µL DMEM άҺ37 5% CO2 չ۲첢Reed-Muench ϡҺϴҺ άҺΪ10μg/mLøDMEM

2

2.1   

ٴɼ أҪΪŻ෢ڳ̺ ɫˮϡ㣨ͼ1AΪʳ˥ˣݣˮţͼ1B෢ʸߴ50% ϣһ㸹к3 4 d  ʱ  ѷĸδк֢״ʼҪΪСͣڱ䱡ͼ1CɫҺ ɫҺ壨ͼ1DС ĤгѪ㣬 ϵĤ ܰ ˮסθ٣֭Ļ ɫҺ塣ʵ  Բ 䡣

ͼ1 ٴ仯

A ɫˮϡ㣻B ݡţ

C Сͱ䱡D Сɫ ɫҺ塣

ͼ1 ٴ  仯

2.2 ԭѧ

СƷRT-PCR ͼ2 ʾ7 СƷ6ݾΪPEDV ԣ85.7%TGEVGAR Ϊԡ

ͼ2 кԭӾͼ

ͼ2 кԭ Ӿͼ

2.3  Ч

2.3.1 ȫ

RT-PCR ǰ ĸPEDV ʾ3  ĸ δĸʳȱ޸кĸʳã޸кʾ  ߰ȫԡ

2.3.2 ˮƽ

ELISA ⿹PEDV IgA ĸƷIgA ΪԣS/P ƽֵΪ0.77    S/P ֱֵΪ0.63 0.62ƷΪPEDV IgGԣ顢 OD ֱֵΪ2.001.84 2.10첻ͼ3

ͼ3 PEDV IgAIgG ˮƽ

ͼ3 PEDV IgAIgG ˮƽ

кͼ4 ʾƽкЧΪ1 351 ƽЧΪ1 188  ƽ кЧΪ1 4.13 P <0.05

ͼ4 кͿЧˮƽ

ͼ4 Чˮƽ

2.3.3  ɼ

۲3 ĸӦк кΪ35.7%кʳϲк ״ã  кأ кΪ55.6% Ӧкݡηѣ

ǵԣҪ7 d Ѿʷ ۸ɼͼ5 ʾ  ߵΪ 飨9.4 ͷ99.2%Ϊ飨9.3 ͷ84.9% 8.3 ͷ77.3% ָ P <0.05

ͼ5 	ƽ 

ͼ5 ƽȽ

3

3.1 ԭ

PEDVTGEVGAR  ԭ ӴԸкٴ֢״仯вѧΪ,ٴ֯ѧϺ֡ ڷʵص顢ʵϣPEDVTGEVGAR RT-PCR ϣʾСƷPEDV ԡTGEV GAR Ϊԣ˵к߲ PEDV

3.2 ߷

ڸóPED ֺ߲ѡʹ  ʵ ƵPEDV ¶ Ʒ ɲͬ߳ ϰ ȫԡ ˮƽϣ ΪPED ߲ ĽУȫãֺ ĸ δ к ӦҲ޸кVero ϸϵ144F ǰʹ ֤ н߰ȫԣ  ֤ ȫԸ֪ˣ3 ߳нϸ߰ȫԡ

ЧҪͨELISA пPEDV IgAIgG 壬к ЧۣʹЧ ȽѾ7 d ʣۺϿPED  ͨڸȾ󣬽СëƤϸƣϸˣëήӪ ϰ к ˮˣ к߻УĤ߾зdzҪáĤڵIgAsIgAСƤϸ  ֵIJԭ̼ĤߣĤѪ IgA 壬ֱӳ 벡ϣԤȾЧ;⣬ ߽ǶȿǣеIgA ĺСıЧ ĸܵPEDV ̼󣬷IgA ϸǨ٣Ӻ֭з IgA 塣ͨ- ;IgA  PEDV ߡ⣬IgM IgG ȣIgA Чֿˮø ⣬IgAкͨ Ⱦ Ҫ塣

У3 ĸΪԣĸ IgA壬  ൱ һ̶֤ۣ2 첻   ЧIgG Ϊָ꣬3 ֮첻ԣԡ3 ĸкƽкЧ 飬ҽ һ  Ҳȫʹ  飬 ˵Чã 硣Ϊ ƣ ЧȷʵЧԽܼľֲĤ ʮ  ޡ⣬PEDV S Ƶ о ֱ꣬ ƷʹCV777 ֮ ϴ죬һ̶ҲӰ ʹЧ ٴ Ч۲ 棬  ĸָккã鳬50% ָкʳ 뿹ˮƽ һ¡к2 4 d ˱ ͳ ĸ 7 d  Ѿʡ7 d   Ϊ飬  ָ  ˵  ṩ

ۺϿĸпPEDV  ˮƽԽߣкͿˮƽԽߣ кԽͣӦ ҲӦԽʹñʵ Ƶ Ʒ ʹ ԽϺã 顣ͨڷ Ȼ ֲ Ĥߣ κ ȫͨ ǻʼǰĹ⣬  ͬʱʾʱעٴPEDV ӦӿPEDV з ٶȡ

ѡԡҵѧ201511 “ҽ”ĿP79-81


Copyright © 2015-2018 利来w666备用ũҵ޹˾ ICP10039185š
֧֣
ַܸ·368
룺510555

erweima