Sohn W M, Nam H W
Department of Parasitology, College of Medicine, Gyeongsang National University, Chinju, Korea.
Korean J Parasitol. 1999 Dec;37(4):249-56. doi: 10.3347/kjp.1999.37.4.249.
A total of 198 sera from stray cats was assayed against Toxoplasma gondii antigen by western blot. Out of 198 sera assayed, 26 sera (13.1%) showed typical blot patterns against T. gondii. When spotted by ELISA absorbance and indirect latex agglutination test (ILAT) titer, all 26 cases were distributed over the cut-off value of ELISA whereas 24 cases (92.3%) were in the positive range of 1:32 or higher and 2 cases in negative range by ILAT. Among western blot negative 172 sera, 162 cases were negative in both ILAT and ELISA while 10 cases were reactive falsely such that three cases were ILAT positive with 1:32 titer and 9 cases were ELISA positive (2 cases overlapped). These 10 cases reacted peculiarly without typical binding pattern in Western blot. Sandwich-ELISA was performed with monoclonal antibodies (mAbs) of Tg563 (30 kDa, SAG1), Tg505 (22 kDa, SAG2), Tg605 (43 kDa, SAG3), Tg556 (28 kDa. GRA2), Tg737 (32 kDa, GRA6), Tg695 (66 kDa, ROP2), Tg786 (42 kDa, ROP6), and Tg621 (32 kDa, anonymous but cytosolic) clone, respectively. All western blot-positive cases were in the positive range and negative cases in the negative range clearly. Among the 10 false reactive cases, 3 cases were in the positive range with one or more mAbs. All mAbs used in this study were confirmed to be specific to T. gondii infection as a standardized sandwich-ELISA to differentiate it from other pathogens.
采用蛋白质印迹法对198份流浪猫血清进行了抗刚地弓形虫抗原检测。在检测的198份血清中，26份血清（13.1%）呈现出针对刚地弓形虫的典型印迹模式。根据酶联免疫吸附测定（ELISA）吸光度和间接乳胶凝集试验（ILAT）滴度进行分析，这26例均分布在ELISA的临界值以上，其中24例（92.3%）ILAT滴度在1:32或更高的阳性范围内，2例为阴性。在蛋白质印迹法检测为阴性的172份血清中，162例ILAT和ELISA均为阴性，10例出现假阳性反应，其中3例ILAT阳性，滴度为1:32，9例ELISA阳性（2例重叠）。这10例在蛋白质印迹法中反应异常，无典型的结合模式。分别用Tg563（30 kDa，SAG1）、Tg505（22 kDa，SAG2）、Tg605（43 kDa，SAG3）、Tg556（28 kDa，GRA2）、Tg737（32 kDa，GRA6）、Tg695（66 kDa，ROP2）、Tg786（42 kDa，ROP6）和Tg621（32 kDa，未知但为胞质蛋白）克隆的单克隆抗体进行夹心ELISA检测。所有蛋白质印迹法阳性病例均明显处于阳性范围，阴性病例处于阴性范围。在10例假阳性反应病例中，3例对一种或多种单克隆抗体呈阳性反应。本研究中使用的所有单克隆抗体均经证实对刚地弓形虫感染具有特异性，作为一种标准化的夹心ELISA可将其与其他病原体区分开来。
