使用固相免疫测定法对商业性溶血巴斯德氏菌疫苗诱导产生的抗体进行比较评估。

Comparative evaluation of antibodies induced by commercial Pasteurella haemolytica vaccines using solid phase immunoassays.

作者信息

Srinand S, Hsuan S L, Yoo H S, Maheswaran S K, Ames T R, Werdin R E

机构信息

Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul 55108, USA.

出版信息

Vet Microbiol. 1996 Apr;49(3-4):181-95. doi: 10.1016/0378-1135(95)00187-5.

DOI:10.1016/0378-1135(95)00187-5

PMID:8734636

Abstract

The objective of this study was to evaluate the ability of four commercial vaccines to elicit antibodies against the leukotoxin (Lkt), capsular polysaccharide (CP), iron regulated outer membrane proteins (IROMPs), and whole cell (WC) antigens of Pasteurella haemolytica A1. Modified double antibody sandwich enzyme linked immunosorbent assays (ELISAs) were developed to measure antibody levels against Lkt, CP and IROMPs. An indirect ELISA was developed to measure the levels of antibody against WC antigens. The ideal cut off points for ELISAs were determined on receiver operating characteristic curves, using sera from 30 calves injected subcutaneously with a live P. haemolytica 12296 strain as positive control and sera from 30 colostrum-deprived calves as negative control. The vaccines evaluated were: 'One Shot' (SmithKline Beecham, West Chester, PA) a bacterin-toxoid, 'Presponse' (Langford Laboratories, Guelph, Ontario) a Lkt-rich culture supermatant, 'Once PMH' (BioCor Inc., Omaha, NE) a modified live vaccine, and 'Septimune' (Fort Dodge laboratories, Fort Dodge, IA) an outer membrane extract. Thirty, 4-6 week old Holstein calves were randomized into 5 groups to receive one of the four vaccines or a placebo (sterile phosphate buffered saline). The calves were vaccinated intramuscularly on day 0 and on day 14, and bled on days, 0, 14, and 28 to measure antibody levels against Lkt, CP, IROMPs, and WC antigens of P. haemolytica Al. 'One Shot', and 'Once PMH' vaccinates showed a significant (P < 0.05) increase in antibody levels against Lkt at 28 days. 'Once PMH' vaccinates also showed significant (P < 0.05) increase in antibody levels against IROMPs at 28 days compared to the other four groups but this increase was not significant over time within the 'Once PMH' group. 'Presponse', 'Once PMH' and 'One Shot' vaccinates showed a significant (P < 0.05) increase in antibody levels against CP over time. These groups also had significantly higher antibody levels against CP, compared to controls and 'Septimune' vaccinates at 14 and 28 days (P < 0.05).

摘要

本研究的目的是评估四种市售疫苗诱导抗溶血巴斯德氏菌A1白细胞毒素（Lkt）、荚膜多糖（CP）、铁调节外膜蛋白（IROMPs）和全细胞（WC）抗原抗体的能力。开发了改良的双抗体夹心酶联免疫吸附测定（ELISA）来检测抗Lkt、CP和IROMPs的抗体水平。开发了一种间接ELISA来检测抗WC抗原的抗体水平。ELISA的理想临界值通过绘制受试者工作特征曲线来确定，使用30头皮下注射活的溶血巴斯德氏菌12296菌株的犊牛血清作为阳性对照，30头初乳缺乏犊牛血清作为阴性对照。评估的疫苗有：“一针灵”（史克必成公司，宾夕法尼亚州韦斯特切斯特），一种菌苗-类毒素；“Presponse”（兰福德实验室，安大略省圭尔夫），一种富含Lkt的培养上清液；“Once PMH”（BioCor公司，内布拉斯加州奥马哈），一种改良活疫苗；以及“Septimune”（福道奇实验室，爱荷华州福道奇），一种外膜提取物。30头4 - 6周龄的荷斯坦犊牛被随机分为5组，分别接受四种疫苗之一或安慰剂（无菌磷酸盐缓冲盐水）。犊牛在第0天和第14天进行肌肉注射，并在第0、14和28天采血，以检测抗溶血巴斯德氏菌A1的Lkt、CP、IROMPs和WC抗原的抗体水平。“一针灵”和“Once PMH”疫苗接种组在第28天时抗Lkt抗体水平显著（P < 0.05）升高。与其他四组相比，“Once PMH”疫苗接种组在第28天时抗IROMPs抗体水平也显著（P < 0.05）升高，但在“Once PMH”组内该升高随时间变化不显著。“Presponse”、“Once PMH”和“一针灵”疫苗接种组抗CP抗体水平随时间显著（P < 0.05）升高。在第14天和第28天时，与对照组和“Septimune”疫苗接种组相比，这些组抗CP抗体水平也显著更高（P < 0.05）。