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设计用于诊断人类囊型棘球蚴病的重组多表位抗原。

Designing a Recombinant Multi-Epitope Antigen of to Diagnose Human Cystic Echinococcosis.

作者信息

Mirzapour Aliyar, Seyyed Tabaei Seyyed Javad, Bandehpour Mojgan, Haghighi Ali, Kazemi Bahram

机构信息

Department of Medical Parasitology and Mycology, International Branch of Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Department of Parasitology and Mycology, School of Medicine, Mashhad Branch, Islamic Azad University, Mashhad, Iran.

出版信息

Iran J Parasitol. 2020 Jan-Mar;15(1):1-10.

Abstract

BACKGROUND

Cystic echinococcosis can cause severe disease and probable death in humans. Epitopes of its antigens play a key role in the sensitivity and specificity of immunodiagnostic tests.

METHODS

Epitope prediction software programs predict the most antigenic linear B-cell epitopes of AgB (8 kD), Ag5, and Ag95. Six such epitopes were predicted and connected by "Gly-Ser" linker and synthesized. The purity of the concentrated recombinant multi-epitope protein was assessed by 15% SDS-PAGE. Overall, 186 serum samples were collected from the Loghman Hakim Hospital and different laboratories, Tehran, Iran, from July 2016 to February 2017. Patients infected with hepatic hydatid cysts, patients infected by other parasites and viruses, and healthy individuals were used to detect the anti-CE IgG using recombinant multi-epitope protein.

RESULTS

Forty-one samples out of 43 cases of hydatidosis were diagnosed correctly as positive, and two were negative. In addition, six negative cases of healthy individual group were diagnosed as positive and negative with rMEP-ELISA and the commercial kit, respectively. Therefore, these six samples were considered as false positive using our method. In addition, a diagnostic sensitivity of 95.3% (95% CI, 84.19% to 99.43%) and a specificity of 95.0% (95% CI, 89.43% to 98.14%) were obtained using optimum cutoff value (0.20). The sensitivity and specificity of the commercial kit was 100%.

CONCLUSION

Our findings showed high diagnostic accuracy of the ELISA test using the developed recombinant protein, which encourages the use of this recombinant multi-epitope protein for rapid serological diagnosis of hydatidosis.

摘要

背景

囊性棘球蚴病可导致人类严重疾病甚至可能死亡。其抗原表位在免疫诊断试验的敏感性和特异性中起关键作用。

方法

利用表位预测软件程序预测AgB(8 kD)、Ag5和Ag95的最具抗原性的线性B细胞表位。预测出6个此类表位,通过“甘氨酸-丝氨酸”接头连接并合成。通过15%十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)评估浓缩重组多表位蛋白的纯度。2016年7月至2017年2月期间,从伊朗德黑兰的洛赫曼·哈基姆医院和不同实验室收集了186份血清样本。使用重组多表位蛋白检测肝包虫囊肿感染患者、其他寄生虫和病毒感染患者以及健康个体的抗囊型包虫病免疫球蛋白G(anti-CE IgG)。

结果

43例包虫病患者中有41例被正确诊断为阳性,2例为阴性。此外,健康个体组的6例阴性病例经重组多表位蛋白酶联免疫吸附测定(rMEP-ELISA)和商用试剂盒分别诊断为阳性和阴性。因此,使用我们的方法将这6个样本视为假阳性。此外,使用最佳临界值(0.20)时,诊断敏感性为95.3%(95%置信区间,84.19%至99.43%),特异性为95.0%(95%置信区间,89.43%至98.14%)。商用试剂盒的敏感性和特异性为100%。

结论

我们的研究结果表明,使用所开发的重组蛋白进行酶联免疫吸附测定(ELISA)试验具有较高的诊断准确性,这促使使用这种重组多表位蛋白进行包虫病的快速血清学诊断。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24e4/7244849/617d83899578/IJPA-15-1-g001.jpg

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