Shen Chenghua, Lee Jong-Ae, Allam Sonia Refaat Ahmed, Bae Young Mee, Han Eun-Taek, Takeo Satoru, Tsuboi Takafumi, Hong Sung-Tae, Choi Min-Ho
Department of Parasitology, Qingdao University Medical College, China.
Diagn Microbiol Infect Dis. 2009 Jul;64(3):334-9. doi: 10.1016/j.diagmicrobio.2009.03.003. Epub 2009 Apr 18.
We evaluated the diagnostic applicability of recombinant proteins from Clonorchis sinensis, the human liver fluke. Four recombinant proteins, 7-kDa protein (Cs7P), 28-kDa cysteine protease (Cs28CP), and 26- and 28-kDa glutathione s-transferases (Cs26GST and Cs28GST), were expressed by wheat germ cell-free protein synthesis system. In ELISA, crude antigen showed the highest sensitivity (92.7%). However, sensitivities of r7P (47.3%), r28CP (30.9%), r26GST (21.8%), and r28GST (14.5%) were dramatically lower. The overall specificities of the crude antigen, r7P, r28CP, r26GST, and r28GST, were 100%, 94.5%, 96.7%, 94.5%, and 98.9%, respectively. Taken together, r7P and r28CP showed moderate sensitivities and high specificities, whereas r26GST and r28GST revealed low sensitivities and high specificities. We demonstrated that recombinant antigens, when used as a single antigen for ELISA, are not sensitive enough to diagnose clonorchiasis. Cocktail or chimeric antigens may be useful to increase the sensitivity of each antigen and may improve the serodiagnosis of clonorchiasis.
我们评估了华支睾吸虫(人体肝吸虫)重组蛋白的诊断适用性。通过小麦胚无细胞蛋白质合成系统表达了四种重组蛋白,即7-kDa蛋白(Cs7P)、28-kDa半胱氨酸蛋白酶(Cs28CP)以及26-kDa和28-kDa谷胱甘肽S-转移酶(Cs26GST和Cs28GST)。在酶联免疫吸附测定(ELISA)中,粗抗原显示出最高的灵敏度(92.7%)。然而,r7P(47.3%)、r28CP(30.9%)、r26GST(21.8%)和r28GST(14.5%)的灵敏度则显著较低。粗抗原、r7P、r28CP、r26GST和r28GST的总体特异性分别为100%、94.5%、96.7%、94.5%和98.9%。总体而言,r7P和r28CP显示出中等灵敏度和高特异性,而r26GST和r28GST则显示出低灵敏度和高特异性。我们证明,重组抗原用作ELISA单一抗原时,对诊断华支睾吸虫病的敏感性不足。混合或嵌合抗原可能有助于提高每种抗原的灵敏度,并可能改善华支睾吸虫病的血清学诊断。