a Wellman Center for Photomedicine, Massachusetts General Hospital , Boston , MA , USA.
b Department of Dermatology , Harvard Medical School , Boston , MA , USA.
Virulence. 2018 Jan 1;9(1):28-63. doi: 10.1080/21505594.2017.1371897. Epub 2017 Dec 8.
Traditional methods of localizing and quantifying the presence of pathogenic microorganisms in living experimental animal models of infections have mostly relied on sacrificing the animals, dissociating the tissue and counting the number of colony forming units. However, the discovery of several varieties of the light producing enzyme, luciferase, and the genetic engineering of bacteria, fungi, parasites and mice to make them emit light, either after administration of the luciferase substrate, or in the case of the bacterial lux operon without any exogenous substrate, has provided a new alternative. Dedicated bioluminescence imaging (BLI) cameras can record the light emitted from living animals in real time allowing non-invasive, longitudinal monitoring of the anatomical location and growth of infectious microorganisms as measured by strength of the BLI signal. BLI technology has been used to follow bacterial infections in traumatic skin wounds and burns, osteomyelitis, infections in intestines, Mycobacterial infections, otitis media, lung infections, biofilm and endodontic infections and meningitis. Fungi that have been engineered to be bioluminescent have been used to study infections caused by yeasts (Candida) and by filamentous fungi. Parasitic infections caused by malaria, Leishmania, trypanosomes and toxoplasma have all been monitored by BLI. Viruses such as vaccinia, herpes simplex, hepatitis B and C and influenza, have been studied using BLI. This rapidly growing technology is expected to continue to provide much useful information, while drastically reducing the numbers of animals needed in experimental studies.
传统的方法来定位和量化致病性微生物在感染的实验动物模型中的存在,大多依赖于牺牲动物,分离组织和计算集落形成单位的数量。然而,几种发光酶(luciferase)的发现,以及细菌、真菌、寄生虫和老鼠的基因工程,使它们在给予荧光素酶底物后发光,或者在细菌 lux 操纵子的情况下,无需任何外源性底物,这提供了一种新的选择。专门的生物发光成像(BLI)相机可以实时记录活的动物发出的光,允许对感染微生物的解剖位置和生长进行非侵入性、纵向监测,其测量方法是 BLI 信号的强度。BLI 技术已用于跟踪创伤性皮肤伤口和烧伤、骨髓炎、肠道感染、分枝杆菌感染、中耳炎、肺部感染、生物膜和牙髓感染和脑膜炎中的细菌感染。经过基因工程改造的发光真菌已被用于研究酵母(念珠菌)和丝状真菌引起的感染。生物发光被用于监测疟疾、利什曼原虫、锥虫和弓形虫引起的寄生虫感染。BLI 还用于研究痘苗病毒、单纯疱疹病毒、乙型肝炎和丙型肝炎病毒以及流感病毒。这项快速发展的技术有望继续提供大量有用的信息,同时大大减少实验研究中所需的动物数量。