动物医学进展
動物醫學進展
동물의학진전
PROGRESS IN VETERINARY MEDICINE
2014年
11期
34-39
,共6页
董林%王艳萍%谢金文%沈志强
董林%王豔萍%謝金文%瀋誌彊
동림%왕염평%사금문%침지강
Balb/c小鼠%动物模型%猪圆环病毒2 型%疫苗效力
Balb/c小鼠%動物模型%豬圓環病毒2 型%疫苗效力
Balb/c소서%동물모형%저원배병독2 형%역묘효력
Balb/c mouse%animal model%Porcine circovirus type 2%vaccine efficacy
为建立以Balb/c小鼠为动物模型的 PCV-2疫苗免疫效力检验方法,选取4周龄 SPF级雌性Balb/c 小鼠20只,随机分成4组(Ⅰ、Ⅱ、Ⅲ、Ⅳ组),每组5只,Ⅰ、Ⅱ和Ⅲ组分别背部皮下1点,背部皮下2点和背部皮下、腿部肌肉2点接种 PCV-2疫苗0.1、0.2、0.2 mL/只,Ⅳ组空白对照组不接种;各免疫组分别于首免疫后7、14、21 d 加强免疫一次,免疫后7、10、14、21、28、35、42 d 采血分离血清,用 ELISA 方法测定PCV-2特异性抗体,确定最佳免疫途径和免疫剂量。另选取4周龄 SPF 级雌性 Balb/c 小鼠,随机分成免疫组、攻毒对照组和健康对照组,免疫组和攻毒对照组于二免后14 d 用 PCV-2强毒株进行攻毒,比较各试验组小鼠在临床症状、病理变化、相对日增重、PCV-2核酸载量等方面差异性,确定小鼠动物模型检测指标。抗体测定表明,一免背部皮下、腿部肌肉2点免疫0.2 mL/只,间隔21 d 加强免疫为最佳免疫程序,免疫后35 d 抗体水平达到峰值,显著高于Ⅰ和Ⅱ组。对照组小鼠免疫攻毒后出现消瘦、被毛凌乱、精神紧张等临床症状,淋巴结肿胀、出血、肺脏出血等病理变化,相对日增重显著降低,PCV-2核酸载量差异在103以上等临床变化。本研究以 Balb/c 小鼠为动物模型建立了 PCV-2疫苗免疫效力检验方法,优化了小鼠免疫途径及剂量,确定了相应的检测指标及技术参数,为 PCV-2疫苗免疫效力检验奠定了基础。
為建立以Balb/c小鼠為動物模型的 PCV-2疫苗免疫效力檢驗方法,選取4週齡 SPF級雌性Balb/c 小鼠20隻,隨機分成4組(Ⅰ、Ⅱ、Ⅲ、Ⅳ組),每組5隻,Ⅰ、Ⅱ和Ⅲ組分彆揹部皮下1點,揹部皮下2點和揹部皮下、腿部肌肉2點接種 PCV-2疫苗0.1、0.2、0.2 mL/隻,Ⅳ組空白對照組不接種;各免疫組分彆于首免疫後7、14、21 d 加彊免疫一次,免疫後7、10、14、21、28、35、42 d 採血分離血清,用 ELISA 方法測定PCV-2特異性抗體,確定最佳免疫途徑和免疫劑量。另選取4週齡 SPF 級雌性 Balb/c 小鼠,隨機分成免疫組、攻毒對照組和健康對照組,免疫組和攻毒對照組于二免後14 d 用 PCV-2彊毒株進行攻毒,比較各試驗組小鼠在臨床癥狀、病理變化、相對日增重、PCV-2覈痠載量等方麵差異性,確定小鼠動物模型檢測指標。抗體測定錶明,一免揹部皮下、腿部肌肉2點免疫0.2 mL/隻,間隔21 d 加彊免疫為最佳免疫程序,免疫後35 d 抗體水平達到峰值,顯著高于Ⅰ和Ⅱ組。對照組小鼠免疫攻毒後齣現消瘦、被毛凌亂、精神緊張等臨床癥狀,淋巴結腫脹、齣血、肺髒齣血等病理變化,相對日增重顯著降低,PCV-2覈痠載量差異在103以上等臨床變化。本研究以 Balb/c 小鼠為動物模型建立瞭 PCV-2疫苗免疫效力檢驗方法,優化瞭小鼠免疫途徑及劑量,確定瞭相應的檢測指標及技術參數,為 PCV-2疫苗免疫效力檢驗奠定瞭基礎。
위건립이Balb/c소서위동물모형적 PCV-2역묘면역효력검험방법,선취4주령 SPF급자성Balb/c 소서20지,수궤분성4조(Ⅰ、Ⅱ、Ⅲ、Ⅳ조),매조5지,Ⅰ、Ⅱ화Ⅲ조분별배부피하1점,배부피하2점화배부피하、퇴부기육2점접충 PCV-2역묘0.1、0.2、0.2 mL/지,Ⅳ조공백대조조불접충;각면역조분별우수면역후7、14、21 d 가강면역일차,면역후7、10、14、21、28、35、42 d 채혈분리혈청,용 ELISA 방법측정PCV-2특이성항체,학정최가면역도경화면역제량。령선취4주령 SPF 급자성 Balb/c 소서,수궤분성면역조、공독대조조화건강대조조,면역조화공독대조조우이면후14 d 용 PCV-2강독주진행공독,비교각시험조소서재림상증상、병리변화、상대일증중、PCV-2핵산재량등방면차이성,학정소서동물모형검측지표。항체측정표명,일면배부피하、퇴부기육2점면역0.2 mL/지,간격21 d 가강면역위최가면역정서,면역후35 d 항체수평체도봉치,현저고우Ⅰ화Ⅱ조。대조조소서면역공독후출현소수、피모릉란、정신긴장등림상증상,림파결종창、출혈、폐장출혈등병리변화,상대일증중현저강저,PCV-2핵산재량차이재103이상등림상변화。본연구이 Balb/c 소서위동물모형건립료 PCV-2역묘면역효력검험방법,우화료소서면역도경급제량,학정료상응적검측지표급기술삼수,위 PCV-2역묘면역효력검험전정료기출。
The animal model of Balb/c mice was established for PCV-2 vaccine immune efficacy in the study. The SPF female Balb/c mice of 4 week old were selected and randomly divided into 4 groups.The mice in-group Ⅰ were immunized with PCV-2 vaccine (0.1 mL/rat)by subcutaneous 1 point.The mice in groupⅡ were immunized with PCV-2 vaccine (0.2 mL/rat)by subcutaneous 2 points.The mice in group Ⅲ were immunized with PCV-2 vaccine (0.2 mL/rat)by subcutaneous 2 points and leg muscles.The mice in groupⅣ were not immunized with PCV-2 vaccine as the blank control group.The mice in immunized groups were immunized secondly in 7,14 and 21 d after first immunity.The blood in mice were took and the sera were separated after 7,10,14,21,28,35,42 days.The PCV-2 specific antibody was detected by ELISA in order to determine the best immune way and immune dose.The 4 week old female SPF Balb/c mice were selected and randomly divided into immune group,challenge control group and healthy control group.The mice in immunized and challenge control groups were attacked challenged by PCV-2 virulent strain after immunization 14 days.The differences of the mice in test groups were compared in clinical symptoms, pathological lesions,relative daily weight gain,PCV-2 DNA load and other aspects in order to determine the detection indexes of the animal model of mice.Mouse ELISA antibody test showed that the best im-mune procedure was subcutaneous and leg muscleinjection (0.2 mL/rat).The peak antibody value ap-peared 35 d after immunization,and was significantly higher than that of group I and II.The control mice showed weight loss,hair messy,mental tension and clinical symptoms,lymph node swelling,bleeding, pulmonary hemorrhage and other pathological lesion after challenge and the relative daily weight gain de-creased significantly.The PCV-2 DNA load difference was more than 103 .This study established the test method for PCV-2 vaccine potency in Balb/c mice as animal model.The dose and route of immunization were optimized,and the detection index and the corresponding technical parameters were determined, which laid a technical support for PCV-2 vaccine potency test.