Le Goff C, Thiaucourt F
PATHOTROP, CIRAD-EMVT Av. d'Agropolis, Montpellier, France.
Vet Microbiol. 1998 Feb 28;60(2-4):179-91. doi: 10.1016/s0378-1135(98)00156-4.
A competitive ELISA, using a specific monoclonal antibody, was designed to detect antibodies to Mycoplasma mycoides subsp. mycoides SC, the agent of contagious bovine pleuropneumonia. One monoclonal antibody was found suitable for such a test, '117/5', it does not cross-react with any of the other mycoplasma species tested, furthermore, its binding is inhibited by positive sera. The cutoff, 50% of inhibition, was determined using a set of negative sera from CBPP-free areas. The sensitivity was controlled with sera from artificially infected animals as well as from sera from areas where CBPP is enzootic. In both cases, cELISA compared favorably with CFT. The precocity of detection was similar but cELISA detected more positives and the positive titers seemed to persist longer than in the case of CFT. Lysis of the antigen used to coat the ELISA plates reduced the variability of fixation and improved the repeatability of the test. A field evaluation is now in progress which will determine the true sensitivity and specificity of the test and also check if antibodies are detected after vaccination.
设计了一种竞争性酶联免疫吸附测定法(ELISA),使用特异性单克隆抗体来检测针对丝状支原体丝状亚种小菌落生物型(Mycoplasma mycoides subsp. mycoides SC)的抗体,该生物型是牛传染性胸膜肺炎的病原体。发现一种单克隆抗体“117/5”适用于该检测,它与所检测的任何其他支原体物种均无交叉反应,此外,其结合受到阳性血清的抑制。使用一组来自无牛传染性胸膜肺炎地区的阴性血清确定了50%抑制率的临界值。用人工感染动物的血清以及来自牛传染性胸膜肺炎地方流行区的血清来控制检测灵敏度。在这两种情况下,酶联免疫吸附测定法(cELISA)与补体结合试验(CFT)相比具有优势。检测的早熟性相似,但cELISA检测出更多阳性结果,并且阳性效价似乎比补体结合试验持续的时间更长。用于包被ELISA板的抗原的裂解减少了固定的变异性并提高了检测的可重复性。目前正在进行现场评估,这将确定该检测的真正灵敏度和特异性,并且还将检查接种疫苗后是否能检测到抗体。