간흡충증에 있어서 자기이동식 ELISA법을 이용한 혈청학적 진단에 관한 연구

초록

Human clonorchiasis is one of the most common trematode diseases and of very important public health problems in Korea. Since skin test always represents neither current infection nor recovery of C. sinensis ova, the immunodiagnosis of human clonorchiasis is not always possible, a more feasible, simple, senstive and specific method is required. Several investigators have described the application of the enzyme-linked immunosorbent assay(ELISA) for the detection of Clonorchis antibodies as an additional serological technique for diagnosing clonorchiasis besides conventional tests. The ELISA method with polycarbonate-coated iron beads as the solid phase and with a magnetic transfer system (M-T ELISA) was proposed in the serodiagnosis of human clonorchiasis. Some of its advantages over the other ELISA systems have been reported as (ⅰ) the uniformity in the surface area of the solid phase involved in the reaction, which is important for the precision of the assay and (ⅱ) the simplicity and economy of simultaneous processing with such a semiautomated system. To investigate the feasibility and diagnostic accuracy of using the M-T ELISA in the sero-diagnosis of human clonorchiasis, sera tested were categorized into 134 clonorchiasis, 10 paragonimiasis, 63 non-infected human cases. To investigate the feasibility of using the standard ELISA with whole blood sample in the sere-diagnosis of clonorchiasis, 26 clonorchiasis subjects were studied. The standard ELISA and the M-T ELISA were performed with Veronal buffered saline antigen of C. sinensis, peroxidase conjugated anti-human IgG and orthophenylenediamine as a substrate by micro-method. The results obtained were as follows: 1. In the preliminary study each optimal dilutions of conjugate and serum by the M-T ELISA were 1/2,000 and 1/100 respectively. 2. The mean 0.D. (optical density) values in the sera of human clonorchiasis were 1.445 ± 0.445 by the standard ELISA and 0.943 ± 0.245 by the M-T ELISA. 3. The mean 0.D. values in the sera of non-infected human cases were 0.292 ± 0.127 by the standard ELISA and 0.308 ± 0.132 by the M-T ELISA. 4. If we proposed the 0.D. value 0.6 as a differential point of positive reaction, the sensitivity of the standard ELISA was 97.0% and of the M-T ELISA was 92.5%. And the specificity of the standard ELISA was 96.8% and of the M-T ELISA was 98.4%. 5. When the results were compared between the standard ELISA and the M-T ELISA of clonorchiasis sera showed that 91.0% gave same reactions. In the clonorchiasis, the O.D. values between the standard ELISA and the M-T ELISA methods were significant at a level of p<0.01. 6. When the results were compared between serum and whole blood sample by the standard ELISA showed same reactions in 95.1%. In the clonorchiasis, the 0.D. value between serum and whole blood sample were high significant at a level of P<0.01. The results between the standard ELISA and the M-T ELISA methods were no differences. So, these findings suggests that the M-T ELISA is more feasible and applicable in the sero-diagnosis of human clonorchiasis in field laboratories.