Vilca-Machaca Lizbeth Sally, Calvay-Sanchez Karen Daphne, Zarate-Sulca Yanina, Jimenez-Vasquez Victor, Ramirez Pablo, Mendoza-Mujica Giovanna
Faculty of Biological Sciences, Universidad Nacional Mayor de San Marcos, Lima 15081, Peru.
Laboratory of Vector-Borne and Zoonotic Bacterial Diseases, National Institute of Health, Lima 15072, Peru.
Pathogens. 2024 Aug 15;13(8):690. doi: 10.3390/pathogens13080690.
Carrion's disease, caused by , is a neglected tropical disease prevalent in the Andean region of South America. Without antimicrobial treatment, this disease has a mortality rate of up to 88% in infected patients. The most common method for diagnosing infection is serological testing. However, the current serological assays are limited in sensitivity and specificity, underscoring the need for the development of novel and more accurate diagnostic tools. Recombinant proteins have emerged as promising candidates to improve the serological diagnosis of Carrion's disease. So, we focused on evaluating the conditions for producing two previously predicted proteins of using the baculovirus-insect cell expression system, mainly the flashBAC ULTRA technology. We assessed various parameters to identify the conditions that yield the highest protein production, including cell lines, temperature, and hours post-infection (hpi). The results showed that the expression conditions for achieving the highest yields of the Prot_689 and Prot_504 proteins were obtained using High Five™ cells at 21 °C and harvesting at 120 hpi. Subsequently, the seroreactivity of recombinant proteins was evaluated using positive sera from patients diagnosed with Carrion's disease. These findings offer valuable insights into the production conditions of recombinant proteins using the baculovirus system, which could significantly contribute to developing more precise diagnostic tools for Carrion's disease. Therefore, this research provides implications for improving diagnostics and potentially developing therapeutic strategies.
卡里翁病由[病原体名称未给出]引起,是一种在南美洲安第斯地区流行的被忽视的热带病。若不进行抗菌治疗,该疾病在感染患者中的死亡率高达88%。诊断[病原体名称未给出]感染最常用的方法是血清学检测。然而,目前的血清学检测在敏感性和特异性方面存在局限性,这凸显了开发新型且更准确诊断工具的必要性。重组蛋白已成为改善卡里翁病血清学诊断的有前景的候选物。因此,我们专注于评估使用杆状病毒 - 昆虫细胞表达系统,主要是flashBAC ULTRA技术生产两种先前预测的蛋白的条件。我们评估了各种参数以确定产生最高蛋白产量的条件,包括细胞系、温度和感染后小时数(hpi)。结果表明,使用High Five™细胞在21°C下培养并在120 hpi收获可获得最高产量的Prot_689和Prot_504蛋白的表达条件。随后,使用来自被诊断为卡里翁病患者的阳性血清评估重组蛋白的血清反应性。这些发现为使用杆状病毒系统生产[病原体名称未给出]重组蛋白的条件提供了有价值的见解,这可能对开发更精确的卡里翁病诊断工具做出重大贡献。因此,本研究对改善诊断以及潜在地开发治疗策略具有启示意义。