禽多杀性巴氏杆菌单克隆抗体的制备及其基本性状研究

将多杀性巴氏杆菌（Ｐ．ｍ．）Ｃ４８－１株（Ｈｅｄｄｌｅｓｔｏｎ１型；Ｃａｒｔｅｒ分型５：Ａ）灭活后全菌免疫的ＢＡＬＢ／Ｃ小鼠脾细胞与ＳＰ／０骨髓瘤细胞在ＰＥＧ１０００作用下融合。用全菌包被的间接ＥＬＩＳＡ方法检测抗体，获得了２３株能稳定分泌抗Ｐ．ｍ．１型单克隆抗体的杂交瘤细胞。杂交瘤细胞培养２个月和冻存３个月后复苏培养，均能稳定分泌特异性单克隆抗体。２３株腹水ＥＬＩＳＡ效价分别从１０－３—１０－１１，无免疫沉淀性，也无凝集性。Ｉｇ类型鉴定表明，３株属于ＩｇＭ，２０株属于ＩｇＧ。间接ＥＬＩＳＡ检测结果表明：２３株单抗只与Ｐ．ｍ．１型起反应，而不与Ｐ．ｍ．３、４、１６型起反应，也不与禽类易感的鸡白痢沙门氏菌、大肠杆菌、葡萄球菌、李氏杆菌起反应，具有型的特异性。用ＩＣ８Ｈ９腹水建立的夹心ＥＬＩＳＡ方法检定Ｐ．ｍ．１型菌株，其敏感性高，特异性强，也更为方便。

Preparation and Identification of Monoclonal Antibodies against Avian Strain of Pasteurella Multocida

stable hybridoma cell lines secreting monoclonal antibodies(McAbs)against serotype 1 (Carter5:A),C48-1, strain of pasteurella multocida, were developed by fusions between the murine myeloma cells SP2/0 and spleen cells of BALB/C mice immunized with C48-1, strain of P.m..Indirect enzyme -linked immunosorbent assays (ELISA)were used to screen those hybridomas producing antibodies.The ELISA titers of McAbs were 10-3 to 10-11.Neither of McAbs have the properties of immunoprecipiation and agglutination. Among them,20 cell lines belong to class IgG and other IgM. All McAbs were only positive against serotype 1,negative against serotype 3,4,16 and the other pathogenic bacteria.The sandwich ELISA estalished with IC3H9 as citic fluid identifies P.m.serotype 1 strain,which is more sensitive,specific and convenient.