Development and evaluation of a colloidal gold immunochromatographic assay based on recombinant protein CatL1D for serodiagnosis of sheep fasciolosis

Abstract

Fasciolosis is a zoonotic parasitic disease that seriously endangers the development of animal husbandry and human health. In order to develop a rapid serological diagnostic method for fasciolosis in ruminants, the CatL1D and CatB4 genes of Fasciola hepatica were amplified by reverse transcription polymerase chain reaction (PCR) and cloned, respectively, and then the CatL-B fusion gene (MeCatL-B) was constructed by gene splicing by overlap extension PCR technique. The recombinant rCatL1D, rCatB4 and rMeCatL-B proteins were then prepared by prokaryotic expression, respectively, and the recombinant protein with high specificity and sensitivity was screened via indirect enzyme-linked immunosorbent assay. Using the selected recombinant protein rCatL1D as a diagnostic antigen, we developed a colloidal gold immunochromatographic assay (CGIA) for detecting F. hepatica-specific antibodies, and 426 serum samples of slaughtered sheep were used to evaluate the sensitivity and specificity of F. hepatica CGIA assay. The results showed that the sensitivity and specificity of rCatL1D protein (100%, 96.67%) were higher than those of rCatB4 (94.29%, 80%) and rMeCatL-B (91.43%, 90%). Compared with the gold standard post-mortem inspection, the specificity and sensitivity of the CGIA method was 100% and 97%, respectively, and the consistency rate between these two methods was 99.3%. These results confirmed that the CGIA method based on rCatL1D protein could be a promising approach for rapid diagnosis of sheep fasciolosis because of its high sensitivity and specificity.

Fasciola gigantica Cathepsin L1H: High Sensitivity and Specificity of Immunochromatographic Strip Test for Antibody Detection

“…In this study, rFgCathL1H was developed to detect antibody anti-FgCathL1H in infected mouse and cattle serum through ICS tests. Additionally, Xifeng et al produced recombinant Cathepsin L1D and developed an ICS test for detecting F. hepatica -specific antibodies [ 34 ]. The Staphylococcal aureus protein A complex was used as a conjugate with gold nanoparticles.…”

“…The T-lines and C-lines were labeled as rCatL1D protein and sheep IgG, respectively. The specificity and sensitivity of the ICS test were 100% and 97%, respectively [ 34 ]. In our study, we used rFgCathL1H conjugated with colloidal gold, as it can be more specific to antibodies than protein A in infected animal serum.…”

Foresight in the diagnosis of trematodiasis: innovations versus routine methods

“…Некоторые исследования изучают фракционированные клетки паразитов, проводя ИФА белков разной массы. При этом довольно быстро достигается воспроизводимый способ получения антигенов с приемлемой чувствительностью и специфичностью [53].…”

Towards the comprehension of fasciolosis (re-)emergence: an integrative overview

“…Different techniques within each type of non-classical detection method have been applied indistinctively in the monitoring of infection within definitive hosts such as ELISAs (Espino et al ., 1998), rapid immunochromatographic tests (Xifeng et al ., 2019), conventional PCR, loop-mediated isothermal amplification (Martínez-Valladares and Rojo-Vázquez, 2016) and high-resolution melting analysis (Moghadamizad et al ., 2020). In addition, detection of snail infections also evolved from the classical parasitological dissection to immunoenzymatic assays (Alba et al ., 2015 a ) and molecular methods (multiplex PCR, Alba et al ., 2015 b ; real-time PCR, Schweizer et al ., 2007; etc.).…”