March J B, Gammack C, Nicholas R
Department of Bacteriology, Moredun Research Institute, Penicuik EH26 0PZ, Scotland.
J Clin Microbiol. 2000 Nov;38(11):4152-9. doi: 10.1128/JCM.38.11.4152-4159.2000.
Latex microspheres (diameter, 8 microm) were coated with anti-Mycoplasma capricolum subsp. capripneumoniae polyclonal immunoglobulin G (IgG) antiserum (anti-F38 biotype). The coated microspheres, when used in a latex agglutination test (LAT), detected M. capricolum subsp. capripneumoniae antigen in the serum of goats with contagious caprine pleuropneumoniae (CCPP). Beads also agglutinated strongly in the presence of purified M. capricolum subsp. capripneumoniae capsular polysaccharide (CPS). Preabsorption of CPS-specific antibodies prior to coating of the beads removed agglutinating activity in the presence of M. capricolum subsp. capripneumoniae, strongly suggesting that CPS is the likely soluble antigen recognized by the test. In addition, the specificity of the LAT exactly mirrored that of an M. capricolum subsp. capripneumoniae CPS-specific monoclonal antibody (WM25): of the 8 other mycoplasma species tested, agglutination was observed only with bovine serogroup 7. The LAT detected all 11 strains of M. capricolum subsp. capripneumoniae examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 1.7 x 10(4) CFU, in a reaction volume of 0.03 ml of serum. With field sera from goats with CCPP, the results of the LAT exhibited a 67% correlation with the results of the currently used complement fixation test (CFT), with the main discrepancy in diagnosis resulting from the increased sensitivity of the LAT compared to that of CFT. This antigen-detection LAT should prove particularly useful in identifying animals in the earliest stages of CCPP and combines sensitivity and low cost with ease of application in the field, without the need for any specialist training or equipment.
将直径8微米的乳胶微球用抗山羊支原体山羊肺炎亚种多克隆免疫球蛋白G(IgG)抗血清(抗F38生物型)包被。当将包被后的微球用于乳胶凝集试验(LAT)时,可检测出患有传染性山羊胸膜肺炎(CCPP)的山羊血清中的山羊支原体山羊肺炎亚种抗原。在纯化的山羊支原体山羊肺炎亚种荚膜多糖(CPS)存在的情况下,微球也会强烈凝集。在包被微球之前对CPS特异性抗体进行预吸附,可消除在山羊支原体山羊肺炎亚种存在时的凝集活性,这有力地表明CPS可能是该检测所识别的可溶性抗原。此外,LAT的特异性与山羊支原体山羊肺炎亚种CPS特异性单克隆抗体(WM25)的特异性完全一致:在所检测的其他8种支原体中,仅与牛血清群7发生凝集。LAT检测了本研究中所检测的所有11株山羊支原体山羊肺炎亚种,在0.03毫升血清的反应体积中,灵敏度水平为2纳克CPS,或相当于1.7×10⁴CFU。对于患有CCPP的山羊的现场血清,LAT的结果与目前使用的补体结合试验(CFT)的结果具有67%的相关性,诊断中的主要差异是由于LAT比CFT的灵敏度更高。这种抗原检测LAT在识别CCPP最早阶段的动物方面应特别有用,并且结合了灵敏度、低成本以及在现场易于应用的特点,无需任何专业培训或设备。