School of Biological Sciences, University of the Punjab, Lahore, Pakistan.
School of Biological Sciences, University of the Punjab, Lahore, Pakistan; Nuffield Department of Medicine, University of Oxford, OX37BN, United Kingdom; School of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan.
Tuberculosis (Edinb). 2022 Sep;136:102253. doi: 10.1016/j.tube.2022.102253. Epub 2022 Aug 30.
Tuberculosis (TB) stays a major cause of death globally after COVID-19 and HIV. An early diagnosis to control TB effectively, needs a fast reliable diagnostic method with high sensitivity. Serodiagnosis involving polyclonal antibodies detection against an antigen of Mycobacterium tuberculosis (Mtb) in serum samples can be instrumental. In our study, Rv3874 and Rv3875 antigens were cloned, expressed, and purified individually and as a chimeric construct in Escherichia coli BL21. Enzyme-Linked Immunosorbent Assay (ELISA) based findings revealed that the Rv3874-Rv3875 chimeric construct was two-fold more sensitive (59.7%) than the individual sensitivities of Rv3874 (28.4%) and Rv3875 (24.9%) for 201 serum TB positive samples. Furthermore, the fusion construct was a little more sensitive (60.4%) for male subjects than that for females (58.8%). Lastly, our preliminary findings, molecular insights of secondary structure, and statistical and in silico analysis of each construct also advocate that CEP can be considered a better immunodiagnostic tool in addition to previously reported EC skin test.
结核病(TB)是继 COVID-19 和 HIV 之后全球主要的死亡原因。为了有效控制结核病,需要一种快速可靠、灵敏度高的诊断方法。血清学诊断通过检测血清样本中针对结核分枝杆菌(Mtb)抗原的多克隆抗体,可以起到辅助作用。在我们的研究中,Rv3874 和 Rv3875 抗原分别在大肠杆菌 BL21 中进行了克隆、表达和纯化,并作为嵌合构建体进行了表达。酶联免疫吸附试验(ELISA)的研究结果表明,与 Rv3874(28.4%)和 Rv3875(24.9%)的个体敏感性相比,Rv3874-Rv3875 嵌合构建体的敏感性高两倍(59.7%),针对 201 份 TB 阳性血清样本。此外,融合构建体对男性的敏感性(60.4%)略高于女性(58.8%)。最后,我们的初步研究结果、二级结构的分子分析以及对每个构建体的统计和计算机分析都表明,CEP 除了之前报道的 EC 皮肤试验外,还可以作为一种更好的免疫诊断工具。