杆状病毒辅助生产巴氏杆菌蛋白:改善卡里昂氏病血清学诊断的潜在策略。

IF 3.3 3区 医学 Q2 MICROBIOLOGY Pathogens Pub Date : 2024-08-15 DOI:10.3390/pathogens13080690
Lizbeth Sally Vilca-Machaca, Karen Daphne Calvay-Sanchez, Yanina Zarate-Sulca, Victor Jimenez-Vasquez, Pablo Ramirez, Giovanna Mendoza-Mujica
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引用次数: 0

摘要

由巴氏杆菌引起的卡里翁病是一种被忽视的热带疾病,流行于南美洲的安第斯地区。如果不进行抗菌治疗,这种疾病感染者的死亡率高达 88%。诊断杆菌感染最常用的方法是血清学检测。然而,目前的血清学检测方法在灵敏度和特异性方面都很有限,因此需要开发新的、更准确的诊断工具。重组蛋白已成为改善卡里昂氏病血清学诊断的有希望的候选方案。因此,我们重点评估了利用杆状病毒-昆虫细胞表达系统(主要是 flashBAC ULTRA 技术)生产两种先前预测的巴氏杆菌蛋白的条件。我们评估了各种参数,以确定产生最高蛋白质产量的条件,包括细胞系、温度和感染后小时数(hpi)。结果显示,Prot_689 和 Prot_504 蛋白产量最高的表达条件是使用 High Five™ 细胞,温度为 21 °C,收获时间为 120 hpi。随后,使用确诊为卡里翁病患者的阳性血清对重组蛋白的血清反应性进行了评估。这些发现为利用杆状病毒系统生产巴氏杆菌重组蛋白的条件提供了有价值的见解,这将大大有助于开发更精确的卡里昂氏病诊断工具。因此,这项研究为改进诊断方法和开发潜在的治疗策略提供了启示。
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Baculovirus-Assisted Production of Bartonella bacilliformis Proteins: A Potential Strategy for Improving Serological Diagnosis of Carrion's Disease.

Carrion's disease, caused by Bartonella bacilliformis, 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 B. bacilliformis 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 B. bacilliformis 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 B. bacilliformis 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.

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来源期刊
Pathogens
Pathogens Medicine-Immunology and Allergy
CiteScore
6.40
自引率
8.10%
发文量
1285
审稿时长
17.75 days
期刊介绍: Pathogens (ISSN 2076-0817) publishes reviews, regular research papers and short notes on all aspects of pathogens and pathogen-host interactions. There is no restriction on the length of the papers. Our aim is to encourage scientists to publish their experimental and theoretical research in as much detail as possible. Full experimental and/or methodical details must be provided for research articles.
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