Detection of Bioluminescent Borrelia burgdorferi from In Vitro Cultivation and During Murine Infection.
Detection of Bioluminescent Borrelia burgdorferi from In Vitro Cultivation and During Murine Infection.
复制标题
体外培养和小鼠感染期间生物发光伯氏疏螺旋体的检测。
DOI：
10.1007/978-1-4939-7383-5_18
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Skare,JonT
中科院分区：
文献类型：
--
作者：
Hyde,JennyA;Skare,JonT
Borrelia burgdorferi, etiologic agent of Lyme disease, is the leading tick-borne disease in the United States with approximately 300,000 cases diagnosed annually. Disease occurs in stages beginning localized infection at the site of a tick bite and progresses to disseminated infection when antibiotic treatment is not administered in a timely manner. A multi-systemic infection develops following dissemination to numerous immunoprotective tissues, such as the heart, bladder, and joints, resulting in late Lyme disease.B. burgdorferiundergoes dynamic genetic regulation throughout mammalian infection and defining the exact role of virulence genes at distinct stages of disease is challenging. The murine model allows for the characterization of the pathogenic function of genes inB. burgdorferi, but traditional end point studies limit the ability to gather data throughout an infection study and greatly increase the required number of mice. Molecular genetic techniques to evaluate and quantitateB. burgdorferiinfection are laborious and costly. To partly circumvent these issues, a codon optimized firefly luciferase, under the control of a constitutive borrelial promoter, was introduced intoB. burgdorferienabling the characterization of mutant or modified strains under in vitro growth conditions and throughout murine infection. The detection of bioluminescentB. burgdorferiis highly sensitive and allows for the repeated real-time quantitative evaluation of borrelial load during murine infection. Furthermore, bioluminescence has also been utilized to evaluate alteration in tissue localization and tissue-specific gene expression ofB. burgdorferi. In this chapter, we describe the generation of bioluminescent borrelial strains along with methods for in vitro, in vivo, and ex vivoB. burgdorferistudies.
