Abstract
BackgroundParagonimiasis is a food-borne trematodiasis, a serious public health issue and a neglected tropical disease. Paragonimus skrjabini is a unique species found in China. Unlike paragonimiasis westermani, it is nearly impossible to make a definitive diagnosis for paragonimiasis skrjabini by finding eggs in sputum or feces. Immunodiagnosis is the best choice to detect paragonimiasis skrjabini. There is an urgent need to develop a novel, rapid and simple immunoassay for large-scale screening patients in endemic areas.Methodology/Principal FindingsTo develop a rapid, simple immunodiagnostic assay for paragonimiasis, rabbit anti-human IgG was conjugated to colloidal gold particles and used to detect antibodies in the sera of paragonimiasis patients. The synthesis and identification of colloidal gold particles and antibody-colloidal gold conjugates were performed. The size of colloidal gold particles was examined using a transmission electron microscope (TEM). The average diameter of colloidal gold particles was 17.46 nm with a range of 14.32–21.80 nm according to the TEM images. The formation of antibody-colloidal gold conjugates was monitored by UV/Vis spectroscopy. Excretory-secretory (ES) antigen of Paragonimus skrjabini was coated on nitrocellulose membrane as the capture line. Recombinant Staphylococcus protein A was used to prepare the control line. This rapid gold immunochromatographic strip was assembled in regular sequence through different accessories sticked on PVC board. The relative sensitivity and specificity of the strip was 94.4% (51/54) and 94.1% (32/34) respectively using ELISA as the standard method. Its stability and reproducibility were quite excellent after storage of the strip at 4°C for 6 months.Conclusions/SignificanceImmunochromatographic strip prepared in this study can be used in a rapid one-step immunochromatographic assay, which is instantaneous and convenient.
Highlights
Paragonimiaisis is a prevalent food-borne zoonosis caused by Paragonimus and is found in African, South American and Asian countries such as China, Japan, Liberia, Nigeria and Venezuela [1,2]
Paragonimiasis skrjabini of humans usually manifests as complex clinical symptoms, which often resulted in misdiagnosis and delayed treatment
Characterization of the colloidal gold particle Colloidal gold particles were synthesized by chemical method using the reduction of chloroauric acid (AuCl3?HCl?4H2O)
Summary
Paragonimiaisis is a prevalent food-borne zoonosis caused by Paragonimus and is found in African, South American and Asian countries such as China, Japan, Liberia, Nigeria and Venezuela [1,2]. Definitive diagnosis of human paragonimiasis is mainly based on the finding of characteristic eggs in sputum or feces. The commonly used tests for paragonimiasis skrjabini were immunodiagnostic methods to detect specific antibodies or antigens using intradermal test (IDT) or Enzymelinked immunosorbent assay (ELISA) [5,6]. There remains a need to develop a novel, rapid and simple immunoassay, especially for screening patients on a large scale in endemic areas. Paragonimus skrjabini is a unique species found in China. It is nearly impossible to make a definitive diagnosis for paragonimiasis skrjabini by finding eggs in sputum or feces. There is an urgent need to develop a novel, rapid and simple immunoassay for large-scale screening patients in endemic areas
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