OBJECTIVE: To establish and evaluate a colloid gold immunochromatographic strip test for the diagnosis of alveolar echinococcosis. METHODS: Total RNA was prepared from Echinococcus multilocularis protoscoleces collected from Xinjiang Uygur Autonomous Region. Em18 gene was obtained by reverse transcription-polymerase chain reaction (RT-PCR). The PCR product was sequenced and cloned into pGEX-3X vector. The recombinant plasmid was expressed and induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) to obtain recombinant protein. The anti-human IgG monoclonal antibodies was conjugated with colloid gold as detecting reagent; the recombinant Em18 antigen and goat anti-mouse IgG were immobilized on nitrocellulose in proper position. The prepared immunochromatographic strip was evaluated using serum samples from patients with alveolar echinococcosis (56), cystic echinococcosis (87), cysticercosis (30), schistosomiasis japonica (10), toxoplasmosis (10) and healthy subjects (50) . Comparison between the immunochromatographic strip test and ELISA was made by kappa statistics. RESULTS: Sensitivity detected by the immunochromatographic strip test was 92.9% (52/56). The cross-reactivity to cystic echinococcosis and cysticercosis was 9.2% (8/87) and 3.3% (1/30), respectively. There was no cross reactivity with schistosomiasis japonica and toxoplasmosis. 4 samples out of 50 healthy people showed false positive reaction. The overall specificity was 93.0 (174/187). Sensitivity and specificity both showed no statistical difference between immunochromatographic strip test and ELISA. High degree of agreement was observed between the strip test and ELISA (kappa = 0.98). CONCLUSION: The developed immunochromatographic strip test using recombinant Em18 antigen as coated antigen is a sensitive, specific, simple and rapid assay for diagnosing alveolar echinococcosis.
OBJECTIVE: To establish and evaluate a colloid gold immunochromatographic strip test for the diagnosis of alveolar echinococcosis. METHODS: Total RNA was prepared from Echinococcus multilocularis protoscoleces collected from Xinjiang Uygur Autonomous Region. Em18 gene was obtained by reverse transcription-polymerase chain reaction (RT-PCR). The PCR product was sequenced and cloned into pGEX-3X vector. The recombinant plasmid was expressed and induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) to obtain recombinant protein. The anti-human IgG monoclonal antibodies was conjugated with colloid gold as detecting reagent; the recombinant Em18 antigen and goat anti-mouse IgG were immobilized on nitrocellulose in proper position. The prepared immunochromatographic strip was evaluated using serum samples from patients with alveolar echinococcosis (56), cystic echinococcosis (87), cysticercosis (30), schistosomiasis japonica (10), toxoplasmosis (10) and healthy subjects (50) . Comparison between the immunochromatographic strip test and ELISA was made by kappa statistics. RESULTS: Sensitivity detected by the immunochromatographic strip test was 92.9% (52/56). The cross-reactivity to cystic echinococcosis and cysticercosis was 9.2% (8/87) and 3.3% (1/30), respectively. There was no cross reactivity with schistosomiasis japonica and toxoplasmosis. 4 samples out of 50 healthy people showed false positive reaction. The overall specificity was 93.0 (174/187). Sensitivity and specificity both showed no statistical difference between immunochromatographic strip test and ELISA. High degree of agreement was observed between the strip test and ELISA (kappa = 0.98). CONCLUSION: The developed immunochromatographic strip test using recombinant Em18 antigen as coated antigen is a sensitive, specific, simple and rapid assay for diagnosing alveolar echinococcosis.