Preparation of Colloidal Gold Immunochromatographic Strip for Detection of Paragonimiasis skrjabini

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.

Diagnosis and surgical management of pericardial effusion due to paragonimiasis

“…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.…”

Teste Imunocromatográfico De Fluxo Lateral: Uma Ferramenta Rápida De Diagnóstico

“…A maioria dos testes rápidos comerciais disponíveis (cerca de 94%) usam partículas de ouro coloidal (KARAKUS & SALIH, 2013), mas podem também ser empregadas no teste: partículas de látex coloridas (GREENWALD et al, 2003), partículas magnéticas (NOR et al, 2012) e moléculas fluorescentes (XIE et al, 2014). As pesquisas acadêmicas que desenvolvem esses testes mostram que o ouro é o mais utilizado (SHYU et al, 2010;DRYGIN et al, 2012;ROCHA, 2012;WIDIYANTI et al, 2013;YONEKITA et al, 2013;VIJAYAN et al, 2013;CHO & IRUDAYARAJ, 2013;WANG et al, 2014), por apresentam vantagens de possuírem superfícies com alta afinidade para proteínas e biomoléculas, o que aumenta os sinais do imunoensaio e permite um fluxo regular através da membrana, além se ser mais estável e fácil de usar (LIU et al, 2011;KAVOSI et al, 2014).…”

“…Dependendo do tipo ou da quantidade dos agentes redutores, há a formação de uma quantidade variável de núcleos centrais. Desta maneira, quanto mais rápida for a redução, uma maior quantidade de núcleos centrais será formada e consequentemente as partículas de ouro coloidal serão menores (HANDLEY, 1989;DISCIPIO, 1996;GORUP, 2010;WANG et al, 2014).…”

“…Várias pesquisas demonstram a utilização tanto de anticorpos monoclonais (DRYGIN et al, 2012;WIDIYANTI et al, 2013;KARAKUS & SALIH, 2013;VIJAYAN et al, 2013;CHO & IRUDAYARAJ, 2013) como os policlonais (DRYGIN et al, 2012;KARAKUS & SALIH, 2013;WANG et al, 2014) para produção desses testes. Tais anticorpos podem ser sensibilizados na linha teste e controle ou serem conjugados com nanopartículas de ouro coloidal.…”

“…As variações de pH para conjugação vão desde pH ácido até pH alcalino, variando entre pH 4,5 até pH 9 (SHYU et al, 2010;WIDIYANTI et al, 2013;WANG et al, 2014;ZANG et al, 2015). A escolha de qual variação de pH estabelecer para a conjugação depende do ponto isoelétrico de cada proteína.…”