Development of an immunodiagnosis method using recombinant PsCP for detection of Paragonimus skrjabini infection in human

Abstract

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.

ELISA based on a recombinant Paragonimus heterotremus protein for serodiagnosis of human paragonimiasis in Thailand

“…In addition to recombinant technology allowing the production of an unlimited amount of antigen, its application in generating a DNA recombinant protein of P. westermani eggs resulted in a significant increase in the sensitivity and specificity of an ELISA for paragonimiasis diagnosis to 90.2 and 100%, respectively, and also useful for an epidemiological study of paragonimiasis [ 18 ]. A recently developed IgG-ELISA using the recombinant cysteine protease antigen (21 kDa) derived from cDNA of the Paragonimus skrjabini juvenile stage was reported to have a sensitivity of 95.5% and no cross-reaction with antibodies from the other human diseases in the study by Yu et al [ 19 ], i.e. echinococcosis granulosus, taeniasis solium, schistosomiasis japonicum and trichinellosis spiralis [ 19 ].…”

“…A recently developed IgG-ELISA using the recombinant cysteine protease antigen (21 kDa) derived from cDNA of the Paragonimus skrjabini juvenile stage was reported to have a sensitivity of 95.5% and no cross-reaction with antibodies from the other human diseases in the study by Yu et al [ 19 ], i.e. echinococcosis granulosus, taeniasis solium, schistosomiasis japonicum and trichinellosis spiralis [ 19 ]. Another recombinant antigen is the 35 kDa cathepsin L lacking signal peptide (rPpsCatL), which is produced from the cDNA of P. pseudoheterotremus worms and belongs to the cysteine protease group.…”

Diagnosis and surgical management of pericardial effusion due to paragonimiasis

“…A specific serologic test for P. skrjabini by using enzyme linked immunosorbent assay (ELISA) was performed in suspected patients. P. skrjabini adults antigens were used in the ELISA with 95.5% sensitivity and 100% specificity (Yu et al, 2017). Cardiac ultrasonography was performed in every patient to detect PE.…”

“…Unlike the other Paragonimus species (ie. P. westermani), it is difficult to find eggs in P. skrjabini infected humans, as humans are not the normal definitive host (Yu et al, 2017;Wang et al, 2014). Furthermore, the immature worm is not usually found in biopsy from patients with larva migrans of P. skrjabini.…”

“…Therefore, the diagnosis in our group is highly suspected by the serologic test and confirmed by pathological examination. Although the ELISA for P. skrjabini in our group (Yu et al, 2017) tested against Schistosoma, Echinococcus, and Trichinella etc, no other species of Paragonimus genus such as P. westermani was tested. So the causative agent in our group is most suggestive of P. skrjabini, however, further investigation is required to confirm the exact species.…”