Savardashtaki Amir, Mostafavi-Pour Zohreh, Arianfar Farzaneh, Sarkari Bahador
Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran.
Recombinant Proteins Laboratory, Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Iran Biomed J. 2019 Jul;23(4):246-52. doi: 10.29252/.23.4.246. Epub 2018 Oct 28.
Cystic echinococcosis (CE) is a helminthic disease caused by the larval form of Echinococcus granulosus. In the present study, the B8/2 subunit of antigen B (AgB) of E. granulosus was expressed in E. coli host and then applied in a diagnostic ELISA set up.
The DNA sequence of AgB8/2 subunit from E. granulosus was extracted from the GenBank and codon-optimized according to E. coli codon usage. The target sequence was cloned in an expression vector (pGEX-4T-1). The produced antigen was used in an ELISA system, and its performance for the diagnosis of human hydatid cyst was evaluated, using sera from CE and non-CE patients, along with the sera from healthy subjects. Moreover, the diagnostic value of the recombinant protein was compared with native AgB, as well as with a commercial kit.
Antibodies to hydatid cyst were detected in 27 out of 30 patients corresponding to a sensitivity of 90% (95% CI: 73-98%). Cross-reaction with sera of non-CE subjects was seen in two cases resulted in a specificity of 93.5% (95% CI: 82-98%) for the test. A sensitivity of 87% and specificity of 90% were found for the native form of the antigen, while the ELISA commercial kit had a sensitivity of 97% and specificity of 95%.
Our data show that rEgAgB8/2 is an appropriate source of antigen for the serological diagnosis of human hydatid cyst. Co-expression of the rEgAgB/2 along with other subunits of AgB may enhance the performances of these antigens for the serodiagnosis of human CE.
囊性棘球蚴病(CE)是由细粒棘球绦虫幼虫引起的一种蠕虫病。在本研究中,细粒棘球绦虫抗原B(AgB)的B8/2亚基在大肠杆菌宿主中表达,然后应用于诊断性酶联免疫吸附测定(ELISA)。
从GenBank中提取细粒棘球绦虫AgB8/2亚基的DNA序列,并根据大肠杆菌密码子使用情况进行密码子优化。将目标序列克隆到表达载体(pGEX-4T-1)中。将产生的抗原用于ELISA系统,使用CE患者、非CE患者以及健康受试者的血清评估其对人包虫囊肿的诊断性能。此外,将重组蛋白的诊断价值与天然AgB以及商用试剂盒进行比较。
30例患者中有27例检测到包虫囊肿抗体,灵敏度为90%(95%置信区间:73-98%)。有2例出现与非CE受试者血清的交叉反应,该检测的特异性为93.�%(95%置信区间:82-98%)。抗原天然形式的灵敏度为87%,特异性为90%,而ELISA商用试剂盒的灵敏度为97%,特异性为95%。
我们的数据表明,重组细粒棘球绦虫AgB8/2是用于人包虫囊肿血清学诊断的合适抗原来源。重组细粒棘球绦虫AgB/2与AgB的其他亚基共表达可能会提高这些抗原对人CE血清学诊断的性能。