Lizbeth Sally Vilca-Machaca, Karen Daphne Calvay-Sanchez, Yanina Zarate-Sulca, Victor Jimenez-Vasquez, Pablo Ramirez, Giovanna Mendoza-Mujica
{"title":"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。","authors":"Lizbeth Sally Vilca-Machaca, Karen Daphne Calvay-Sanchez, Yanina Zarate-Sulca, Victor Jimenez-Vasquez, Pablo Ramirez, Giovanna Mendoza-Mujica","doi":"10.3390/pathogens13080690","DOIUrl":null,"url":null,"abstract":"<p><p>Carrion's disease, caused by <i>Bartonella bacilliformis</i>, 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 <i>B. bacilliformis</i> 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 <i>B. bacilliformis</i> 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 <i>B. bacilliformis</i> 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.</p>","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.3000,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11357310/pdf/","citationCount":"0","resultStr":"{\"title\":\"Baculovirus-Assisted Production of <i>Bartonella bacilliformis</i> Proteins: A Potential Strategy for Improving Serological Diagnosis of Carrion's Disease.\",\"authors\":\"Lizbeth Sally Vilca-Machaca, Karen Daphne Calvay-Sanchez, Yanina Zarate-Sulca, Victor Jimenez-Vasquez, Pablo Ramirez, Giovanna Mendoza-Mujica\",\"doi\":\"10.3390/pathogens13080690\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Carrion's disease, caused by <i>Bartonella bacilliformis</i>, 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 <i>B. bacilliformis</i> 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 <i>B. bacilliformis</i> 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 <i>B. bacilliformis</i> 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.</p>\",\"PeriodicalId\":19758,\"journal\":{\"name\":\"Pathogens\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.3000,\"publicationDate\":\"2024-08-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11357310/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Pathogens\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3390/pathogens13080690\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Pathogens","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3390/pathogens13080690","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
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.
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.