应用天然组织蛋白酶-L 蛋白酶和重组组织蛋白酶 L1-D 对水牛片形吸虫感染的血清学诊断
Serodiagnosis of Fasciola gigantica Infection in Buffaloes with Native Cathepsin-L Proteases and Recombinant Cathepsin L1-D.
机构信息
Sam Higginbottom University of Agriculture, Technology and Sciences, Prayagraj, U.P., 211007, India.
ICAR-Indian Veterinary Research Institute, Izatnagar, U.P., 243122, India.
出版信息
Acta Parasitol. 2020 Jun;65(2):413-421. doi: 10.2478/s11686-020-00177-3. Epub 2020 Feb 19.
AIM
Serodiagnosis of Fasciola gigantica natural infection in buffaloes with recombinant cathepsin L1-D and native cathepsin-L protease antigens.
METHODS
The recombinant cat L1-D antigen was expressed in prokaryotic expression system and native cathepsin-L proteases were purified by alcoholic fractionation from adult F. gigantica flukes. Buffaloes (n = 325) were screened for anti-Fasciola antibodies with the above antigens in immunoglobulin-G-enzyme linked immunosorbent assay (IgG-ELISA).
RESULTS
The recombinant cat L1-D antigen showed positive reactivity with 101/122 necropsy positive animals but 21/122 necropsy confirmed positive animals were negative in this ELISA (sensitivity 82.8%). However, 30/203 (14.8%) necropsy negative animals for Fasciola were seropositive with specificity of 85.2%. With native cat-L protease, 104/122 necropsy confirmed positive animals were ELISA positive but 18/122 necropsy positive animals were seronegative, thereby depicting the sensitivity of 85.2%. But ELISA with this antigen showed 27/203 (13.3%) necropsy negative animals as positive (specificity 86.7%).
CONCLUSIONS
Comparative evaluation of both the antigens showed that they are suitable for serodiagnosis of F. gigantica infection in buffalo herds.
目的
用重组 cathepsin L1-D 和天然 cathepsin-L 蛋白酶抗原检测水牛自然感染华支睾吸虫的血清学诊断。
方法
在原核表达系统中表达重组 cat L1-D 抗原，并用酒精分级分离法从成虫 F. gigantica 中纯化天然 cathepsin-L 蛋白酶。用上述抗原的免疫球蛋白 G-酶联免疫吸附试验（IgG-ELISA）检测水牛（n = 325）的抗 Fasciola 抗体。
结果
重组 cat L1-D 抗原与 122 例剖检阳性动物中的 101/122 例呈阳性反应，但在该 ELISA 中，21/122 例剖检确认阳性动物为阴性（敏感性 82.8%）。然而，30/203（14.8%）例剖检为 Fasciola 阴性的动物呈血清阳性，特异性为 85.2%。用天然 cat-L 蛋白酶，122 例剖检确认阳性动物中的 104/122 例 ELISA 阳性，但 18/122 例剖检阳性动物为血清阴性，因此敏感性为 85.2%。但是，该抗原的 ELISA 显示 203 例剖检阴性动物中的 27/203（13.3%）为阳性（特异性 86.7%）。
结论
两种抗原的比较评估表明，它们适用于水牛群华支睾吸虫感染的血清学诊断。
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