Yu Shasha, Zhang Xilin, Chen Wenbi, Zheng Hong, Ai Guoping, Ye Nan, Wang Ying
Institute of Tropical Medicine, Third Military Medical University, Chongqing, 400038, China.
Department of Pathogen Biology, Third Military Medical University, Chongqing, 400038, China.
Parasitol Res. 2017 Jan;116(1):377-385. doi: 10.1007/s00436-016-5300-2. Epub 2016 Oct 28.
Paragonimiasis skrjabini is a kind of zoonosis and prevalent in 16 provinces in China, such as Chongqing, Fujian, Sichuan, and Yunnan. However, sensitive and efficient diagnostic methods for the infection are limited. In order to provide a more convenient and simple method for serologic diagnosis, the recombinant P. skrjabini cysteine protease (PsCP) was expressed, purified, and then used to develop an indirect enzyme-linked immunosorbent assay (ELISA) for detecting anti-PsCP antibodies in human. Given the positive/negative cutoff value as 0.606, the maximum dilution of human sera in which anti-PsCP antibodies could be detected was 1:12,800. In addition, the coefficients of variation (CVs) of inter-assay and intra-assay experiments were both below 10 %. Furthermore, the sensitivity of the PsCP-based ELISA was 95.5 %, and the indirect ELISA displays no cross-reactivity with human antisera against Echinococcus granulosus, Taenia solium, Schistosoma japonicum, and Trichinella spiralis, either. In conclusion, recombinant PsCP was readily produced and used to establish a simple PsCP-based ELISA that provided a highly specific and sensitive method for analysis of clinical samples. Besides, the method can also probably be used to diagnose P. skrjabini infection in animals.
斯氏肺吸虫病是一种人畜共患病，在中国的16个省份流行，如重庆、福建、四川和云南。然而，针对该感染的灵敏且高效的诊断方法有限。为了提供一种更便捷、简单的血清学诊断方法，表达并纯化了重组斯氏肺吸虫半胱氨酸蛋白酶（PsCP），然后用其开发了一种间接酶联免疫吸附测定（ELISA），用于检测人体中的抗PsCP抗体。以0.606作为阳性/阴性临界值，能够检测到抗PsCP抗体的人血清最大稀释度为1:12,800。此外，批间和批内实验的变异系数（CV）均低于10%。而且，基于PsCP的ELISA灵敏度为95.5%，间接ELISA与人体针对细粒棘球绦虫、猪带绦虫、日本血吸虫和旋毛虫的抗血清也无交叉反应。总之，重组PsCP易于制备，并用于建立一种简单的基于PsCP的ELISA，该方法为临床样本分析提供了一种高度特异且灵敏的方法。此外，该方法或许还可用于诊断动物的斯氏肺吸虫感染。
