Saadat Saman, Sajadi Mohammad M, Alikhani Mohammad Yousef, Rikhtegaran Tehrani Zahra, Yousefi Mashouf Rasoul
Department of Microbiology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran; Institute of Human Virology at the University of Maryland, School of Medicine, MD, USA.
Institute of Human Virology at the University of Maryland, School of Medicine, MD, USA.
J Microbiol Methods. 2018 Jan;144:186-191. doi: 10.1016/j.mimet.2017.12.001. Epub 2017 Dec 5.
Mycoplasma hominis is an opportunistic pathogen of the human genital tract. Detection of antibodies against this organism in human serum or plasma is theoretically unreliable because of high variation in bacterial surface antigens. In this study, we applied the bioinformatics tools to design a chimeric protein constructed of specific, conserved and predicted immuno-dominant epitopes from two different membrane proteins, P120 and P80.
Linear B-cell epitopes of P120 and P80 were predicted and evaluated by bioinformatics tools and the designed chimeric protein was expressed in Escherichia coli. The chimeric protein, Mh128, was further analyzed in terms of immuno-reactivity by western blotting and enzyme immuno-sorbent assay (ELISA).
We found eight specific, conserved and immuno-dominant epitopes within P120 and P80 based on the bioinformatic studies. The constructed chimeric protein showed immuno-reaction in both western-blotting and ELISA tests.
Because of extensive variation of genomic and antigenic structure, diagnosis of M. hominis infection is difficult. Mh128 as a predicted specific and conserved recombinant protein can be potentially used for sero-diagnosis of M. hominis infection. We plan to develop an immuno-assay based on Mh128 and further evaluate the clinical specificity and sensitivity of the method.
人型支原体是人类生殖道的一种机会致病菌。由于细菌表面抗原高度变异,在人血清或血浆中检测针对该病原体的抗体在理论上并不可靠。在本研究中,我们应用生物信息学工具设计了一种嵌合蛋白,该蛋白由来自两种不同膜蛋白P120和P80的特异性、保守且预测为免疫显性的表位构建而成。
通过生物信息学工具预测并评估P120和P80的线性B细胞表位,并在大肠杆菌中表达设计的嵌合蛋白。通过蛋白质印迹法和酶联免疫吸附测定(ELISA)进一步分析嵌合蛋白Mh128的免疫反应性。
基于生物信息学研究,我们在P120和P80中发现了8个特异性、保守且免疫显性的表位。构建的嵌合蛋白在蛋白质印迹法和ELISA检测中均显示出免疫反应。
由于基因组和抗原结构的广泛变异,人型支原体感染的诊断较为困难。Mh128作为一种预测的特异性和保守重组蛋白,有可能用于人型支原体感染的血清学诊断。我们计划开发一种基于Mh128的免疫测定方法,并进一步评估该方法的临床特异性和敏感性。