Kashyap Rajpal S, Bhullar Shradha S, Chandak Nitin H, Taori Girdhar M
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur, 440010, India.
Methods Mol Biol. 2016;1426:39-50. doi: 10.1007/978-1-4939-3618-2_4.
Synthetic peptide-based diagnosis of Chikungunya can be an efficient and more accessible approach in immunodiagnostics. Here, we describe the identification of Chikungunya-specific 40 kD protein for development of synthetic peptide-based enzyme-linked immunosorbent assay for the detection of Chikungunya virus-specific antibodies in the patient's sample. The total sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profile of the patient's sample can be done to identify specific protein bands. The identified proteins can be subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) for characterization. After characterization, immunogenic peptides can be designed using softwares and subsequently synthesized chemically. The peptides can be used to develop more specific, sensitive, and simpler diagnostic assay.
基于合成肽的基孔肯雅热诊断在免疫诊断中可能是一种高效且更易实现的方法。在此,我们描述了用于开发基于合成肽的酶联免疫吸附测定以检测患者样本中基孔肯雅病毒特异性抗体的基孔肯雅热特异性40 kD蛋白的鉴定。可对患者样本进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳总蛋白谱分析以鉴定特定蛋白条带。鉴定出的蛋白可进行液相色谱-串联质谱(LC-MS/MS)分析以进行表征。表征后,可使用软件设计免疫原性肽,随后进行化学合成。这些肽可用于开发更特异、灵敏且更简单的诊断测定。