背景:Opisthorchiasis和胆管癌(CCA)仍然是许多东南亚国家的公共卫生问题。尽管opistorchiasis的患病率正在下降,报告的病例倾向于轻度感染。因此,使用灵敏的方法进行早期检测是必要的。已经开发了几种灵敏的方法来检测opisthorchiasis。抗原蛋白的免疫检测已被提出作为检查opisthorchiasis的灵敏方法。
方法:Opisthorchisviverrini抗原蛋白,包括组织蛋白酶B(OvCB),天冬酰胺酰内肽酶(OvAEP),和组织蛋白酶F(OvCF),用于构建多抗原蛋白。OvCB的蛋白质序列,OvAEP,OvCF,B细胞表位的概率很高,是使用BeipPred1.0和IEDB分析资源选择的。这些蛋白片段结合形成OvCB_OvAEP_OvCF重组DNA,然后用于在大肠杆菌菌株BL21(DE3)中生产重组蛋白。使用免疫印迹法评估了重组蛋白作为opisthorchiasis病诊断靶标的效力,并与金标准方法进行了比较。改进的福尔马林-醚浓缩技术。
结果:重组OvCB_OvAEP_OvCF蛋白与总免疫球蛋白G(IgG)抗体在流行地区显示出针对光强度O.viverrini感染的强反应性。因此,据报道,诊断opisthorchiasis的敏感性很高(100%)。然而,与来自其他蠕虫和原生动物感染的血清的交叉反应性(包括头虫病,圆线虫病,贾第鞭毛虫病,大肠杆菌感染,肠病,和棘皮组织混合感染。和牛带虫属物种。)并且与非寄生虫感染患者的血清没有反应,导致特异性降低了78.4%。此外,假阴性率(FNR),假阳性率(FPR),阳性预测值(PPV),负预测值(NPV),诊断准确率为0%,21.6%,81.4%,100%,88.9%,分别。
结论:重组OvCB_OvAEP_OvCF蛋白在检测opisthorchiasis方面的高灵敏度表明其作为opisthorchiasis筛查靶标的潜力。尽管如此,减少交叉反应性的研究应通过检测其他样品类型中的其他抗体来进行,比如唾液,尿液,还有粪便.
BACKGROUND: Opisthorchiasis and cholangiocarcinoma (CCA) continue to be public health concerns in many Southeast Asian countries. Although the prevalence of opisthorchiasis is declining, reported cases tend to have a light-intensity infection. Therefore, early detection by using sensitive methods is necessary. Several sensitive methods have been developed to detect opisthorchiasis. The immunological detection of antigenic proteins has been proposed as a sensitive method for examining opisthorchiasis.
METHODS: The Opisthorchis viverrini antigenic proteins, including cathepsin B (OvCB), asparaginyl endopeptidase (OvAEP), and cathepsin F (OvCF), were used to construct multi-antigenic proteins. The protein sequences of OvCB, OvAEP, and OvCF, with a high probability of B cell epitopes, were selected using BepiPred 1.0 and the IEDB Analysis Resource. These protein fragments were combined to form OvCB_OvAEP_OvCF recombinant DNA, which was then used to produce a recombinant protein in Escherichia coli strain BL21(DE3). The potency of the recombinant protein as a diagnostic target for opisthorchiasis was assessed using
immunoblotting and compared with that of the gold standard method, the modified formalin-ether concentration technique.
RESULTS: The recombinant OvCB_OvAEP_OvCF protein showed strong reactivity with total immunoglobulin G (IgG) antibodies against light-intensity O. viverrini infections in the endemic areas. Consequently, a high sensitivity (100%) for diagnosing opisthorchiasis was reported. However, cross-reactivity with sera from other helminth and protozoan infections (including taeniasis, strongyloidiasis, giardiasis, E. coli infection, enterobiasis, and mixed infection of Echinostome spp. and Taenia spp.) and no reactivity with sera from patients with non-parasitic infections led to a reduced specificity of 78.4%. In addition, the false negative rate (FNR), false positive rate (FPR), positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy were 0%, 21.6%, 81.4%, 100%, and 88.9%, respectively.
CONCLUSIONS: The high sensitivity of the recombinant OvCB_OvAEP_OvCF protein in detecting opisthorchiasis demonstrates its potential as an opisthorchiasis screening target. Nonetheless, research on reducing cross-reactivity should be undertaken by detecting other antibodies in other sample types, such as saliva, urine, and feces.