Wang Jinhui, He Kangxin, Wu Zhengjiao, Jin Weikun, Wu Wende, Guo Yanfeng, Zhang Weiyu, Di Wenda
College of Animal Science and Technology, Guangxi University, Nanning, China.
Guangxi Zhuang Autonomous Region Engineering Research Center of Veterinary Biologics, Guangxi University, Nanning, China.
Front Vet Sci. 2022 Sep 16;9:1004932. doi: 10.3389/fvets.2022.1004932. eCollection 2022.
, a tropical liver fluke, infects buffalo in Asian and African countries, causing significant economic losses and posing public health threats. The diagnostic of buffalo fascioliasis caused by is vital in fascioliasis control and preventation. The 22nd gel filtration chromatography fraction of Excretory-Secretory Products (ESP), namely 22 (F22), which was used as a diagnostic antigen in indirect ELISA, has demonstrated great potential for fascioliasis diagnosing. In the absence of rapid diagnostic methods, the use of a colloidal gold immunochromatographic strip based on F22 was applied to detect infection in buffalo.
In the present study, the 22 gel filtration chromatography fraction of ESP (F22) was used as an antigen to establish the colloidal gold-based immunochromatographic strip (ICS). The nitrocellulose membrane was incubated with F22 at the test line (T line) and goat anti-mouse secondary antibody at the control line (C line). The mouse anti-buffalo secondary antibody 2G7 conjugated to colloidal gold particles was used as the detection system for line visualization. The strip was assembled and developed by optimizing reaction conditions. The sensitivity, specificity, stability, and early diagnostic value of the strip were evaluated employing buffalo-derived sera.
An immunochromatographic strip for the rapid detection of antibodies against -ICS was developed. The strip demonstrated high sensitivity and specificity. Sensitivity tests confirmed positive results even when the positive reference serum was diluted 4,096 times. Except for one -positive serum that tested positive via ICS, specificity tests confirmed no cross-reactivity with other positive sera of and . The strip remained stable after storage at 4°C for up to 3 months. In infected buffalo, antibodies could be detected as early as 14-21 days post-infection. The detection of 17 positive sera yielded an 82.4% positive rate via ICS vs. a 100.0% positive rate via ELISA based on ESP. For ICS, the 95% confidence interval of sensitivity was 84.8-95.4%, while specificity was 4.2-14.7%.
The immunochromatographic strip ICS developed in this study provides a simple and rapid method of antibody detection and infected buffalo monitoring in the field.
[肝片吸虫名称]，一种热带肝吸虫，在亚洲和非洲国家感染水牛，造成重大经济损失并构成公共卫生威胁。诊断由[肝片吸虫名称]引起的水牛肝片吸虫病对于肝片吸虫病的控制和预防至关重要。[肝片吸虫名称]排泄分泌产物（ESP）的第22个凝胶过滤层析组分，即[肝片吸虫名称]22（F22），在间接ELISA中用作诊断抗原，已显示出在肝片吸虫病诊断方面的巨大潜力。在缺乏快速诊断方法的情况下，基于F22的胶体金免疫层析试纸条被用于检测水牛中的[肝片吸虫名称]感染。
在本研究中，ESP的第22个凝胶过滤层析组分（F22）用作抗原，建立基于胶体金的免疫层析试纸条（ICS）。将硝酸纤维素膜在检测线（T线）用F22孵育，在对照线（C线）用山羊抗小鼠二抗孵育。与胶体金颗粒偶联的小鼠抗水牛二抗2G7用作条带可视化的检测系统。通过优化反应条件组装并开发试纸条。使用水牛来源的血清评估试纸条的敏感性、特异性、稳定性和早期诊断价值。
开发了一种用于快速检测抗[肝片吸虫名称] - ICS抗体的免疫层析试纸条。该试纸条显示出高敏感性和特异性。敏感性测试证实，即使阳性参考血清稀释4096倍仍呈阳性结果。除一份[肝片吸虫名称]阳性血清通过ICS检测为阳性外，特异性测试证实与其他[肝片吸虫名称]和[另一种寄生虫名称]阳性血清无交叉反应。试纸条在4°C储存长达3个月后仍保持稳定。在感染的水牛中，感染后14 - 21天即可检测到抗体。检测17份阳性血清，通过ICS的阳性率为82.4%，而基于ESP的ELISA阳性率为100.0%。对于ICS，敏感性的95%置信区间为84.8 - 95.4%，特异性为4.2 - 14.7%。
本研究开发的免疫层析试纸条ICS提供了一种在现场简单快速检测[肝片吸虫名称]抗体和监测感染水牛的方法。
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