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目的探讨重组日本血吸虫琥珀酸脱氢酶铁硫蛋白(rSjSDISP)用于血吸虫病的诊断价值。方法利用经原核表达并纯化的rSjSDISP,建立基于重组抗原的快速酶联免疫吸附试验(F-ELISA),检测血吸虫感染不同时期的兔血清、健康人血清、寄生虫病患者血清(其中包括血吸虫病患者血清、肝吸虫病患者血清、肺吸虫病患者血清),比较rSjSDISP F-ELISA与SEA-IHA两者的早期诊断价值及对rSjSDISP F-ELISA诊断方法的诊断效能做出评价;并与SEA-IHA法和SEA FELISA法平行检测血吸虫病疫区335份待测人血清比较3者的敏感性和特异性。结果成功构建pET32a/rSjSDISP重组质粒,当IPTG终浓度为0.1mmol/L,28℃诱导6h时SjSDISP重组蛋白表达量达到最大。rSjSDISP F-ELISA与SEA-IHA可区别出血吸虫感染28d后的兔血清,两者检测结果差异无统计学意义;rSjSDISP F-ELISA敏感度为94.4,特异度为93.0,Youden指数为0.88。在诊断335份血吸虫病疫区现场血清中,该方法与SEA-IHA、SEA F-ELISA具有相似的敏感性和特异性(P>0.05)。结论 rSjSDISP重组抗原可作为一种成分均一的优势诊断抗原,替代成分复杂的虫卵可溶性抗原(SEA)用于血吸虫病的诊断。rSjSDISP F-ELISA方法操作简便、快速,抗原制备易于标化,稳定性强,在血吸虫病诊断上具有较大的应用潜能。
Objective To investigate the diagnostic value of recombinant Schistosoma japonicum succinate dehydrogenase iron-sulfur protein (rSjSDISP) for schistosomiasis. Methods A rapid enzyme-linked immunosorbent assay (F-ELISA) based on rSjSDISP was used to detect the sera of schistosomiasis infected rabbits at different periods, serum of healthy people and serum of parasitic patients (including schistosomiasis Serum of patients, patients with paragonimiasis and paragonimiasis patients). The diagnostic value of both rSjSDISP F-ELISA and SEA-IHA was compared with that of rSjSDISP F-ELISA. IHA method and the SEA FISA method parallel detection of schistosomiasis prevalence area of 335 human serum to be tested compared three sensitivity and specificity. Results The recombinant plasmid pET32a / rSjSDISP was successfully constructed. When the final concentration of IPTG was 0.1mmol / L, the expression of SjSDISP recombinant protein was maximized when induced by 28 ℃ for 6 hours. The results of rSjSDISP F-ELISA and SEA-IHA showed that there was no significant difference between the two groups. The sensitivity of rSjSDISP F-ELISA was 94.4, the specificity was 93.0, and the Youden index was 0.88. This method is similar to SEA-IHA and SEA F-ELISA in the diagnosis of sera from 335 schistosomiasis endemic areas (P> 0.05). Conclusion The rSjSDISP recombinant antigen can be used as a predominant diagnostic antigen for the composition, instead of the complicated complex soluble antigen (SEA) for the diagnosis of schistosomiasis. The rSjSDISP F-ELISA method is simple, rapid, easy to standardize antigen preparation, strong stability, and has great potential in the diagnosis of schistosomiasis.