Zhou Yan, Xu Xue-Nian, Yao Kai-Ling, Zhang Hong-Man, Cheng Na, Bao Yi-Fang, Zhang Lu-Juan, Xu Bin, Jiang He, Li Xue-Ming, Chun Peter, Feng Zheng
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai 200025, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2011 Jun;29(3):172-6.
To develop and preliminarily evaluate two immunodiagnostic methods for clonorchiasis using Clonorchis sinensis PPMP I antigen Cs2 recombinant protein (rCs2).
Using the soluble rCs2, an indirect ELISA and a colloidal-gold immuno-chromatography assay (GICA) dynamic flow strip was developed for detecting specific antibodies in serum. Serum samples from 35 egg-positive clonorchiasis patients, 33 healthy individuals, 15 schistosomiasis patients, 15 paragonimiasis westermani patients and 13 cysticercosis patients were examined by ELISA and GICA strip test. To further evaluate the diagnostic value of these two methods, eight New Zealand rabbits were randomly divided into infected group and treatment group. Each rabbit was infected with 600 C. sinensis metacercaria. Rabbits in treatment group were treated with praziquantel [150 mg/(kg x d) x 2d] individually at day 56 post-infection. ELISA and GICA strip test were used to observe the dynamic changes of specific antibodies against rCs2 in the two parallel groups during the period of 0-44 weeks.
The sensitivity, specificity and total coincidence rate determined by the ELISA method were 71.4% (25/35), 93.4% (71/76), and 86.5% (96/111), respectively, and the cross reaction with schistosomiasis, paragonimiasis and cysticercosis patients were 1/15, 1/15, and 1/13, respectively. The sensitivity, specificity and coincidence rate in the GICA strip test were 85.7% (30/35), 92.1% (70/76), and 90.1%(100/111), respectively. In C sinensis infected rabbits, antibodies level began to increase at 4 weeks after infection, peaked at the 6th week, and declined rapidly to a lower level in the 20th week, while the changing pattern of antibodies level in the treatment group was similar with that of infected group (P > 0.05). In the GICA strip test, antibodies in two groups could be detected in 4-16 weeks.
Indirect ELISA and the GICA dynamic flow strip developed in this study may be of value in the immunodiagnosis of clonorchiasis.
利用华支睾吸虫PPMP I抗原Cs2重组蛋白（rCs2）开发并初步评估两种华支睾吸虫病免疫诊断方法。
使用可溶性rCs2，开发间接酶联免疫吸附测定（ELISA）和胶体金免疫层析分析（GICA）动态流动试纸条用于检测血清中的特异性抗体。用ELISA和GICA试纸条检测法对35例华支睾吸虫病虫卵阳性患者、33例健康个体、15例血吸虫病患者、15例卫氏并殖吸虫病患者和13例囊尾蚴病患者的血清样本进行检测。为进一步评估这两种方法的诊断价值，将8只新西兰兔随机分为感染组和治疗组。每只兔感染600个华支睾吸虫囊蚴。治疗组兔在感染后第56天分别用吡喹酮[150 mg/(kg·d)×2d]进行治疗。采用ELISA和GICA试纸条检测法观察0~44周期间两组平行实验中针对rCs2的特异性抗体动态变化。
ELISA法测定的敏感性、特异性和总符合率分别为71.4%（25/35）、93.4%（71/76）和86.5%（96/111），与血吸虫病、卫氏并殖吸虫病和囊尾蚴病患者的交叉反应分别为1/15、1/15和1/13。GICA试纸条检测法的敏感性、特异性和符合率分别为85.7%（30/35）、92.1%（70/76）和90.1%（100/111）。在感染华支睾吸虫的兔中，抗体水平在感染后第4周开始升高，第6周达到峰值，在第20周迅速下降至较低水平，而治疗组抗体水平的变化模式与感染组相似（P>0.05）。在GICA试纸条检测法中，两组在4~16周均可检测到抗体。
本研究开发的间接ELISA和GICA动态流动试纸条可能在华支睾吸虫病的免疫诊断中具有价值。
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