The development of FastMyco⢠: A novel isothermal colorimetric assay for the rapid detection of mycoplasma contamination .
FastMyco™ 的开发:一种用于快速检测支原体污染的新型等温比色测定法。
基本信息
- 批准号:10080974
- 负责人:
- 金额:$ 27.43万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2020
- 资助国家:美国
- 起止时间:2020-09-01 至 2022-02-28
- 项目状态:已结题
- 来源:
- 关键词:Acholeplasma laidlawiiAddressAdoptedAgricultureAlkaliesArginineBindingBiologicalBiological AssayBiomedical ResearchCell Culture TechniquesCell LineCell MaintenanceCell WallCell membraneColorCoupledCulture MediaCytolysisDNADecision MakingDetectionDetergentsDevelopmentDiagnostic testsDyesEnzymesEquipmentExhibitsFluorescenceFreeze DryingGentian VioletHandHumanIncubatorsIndividualInfrastructureLaboratoriesLinkMagnesiumMajor GrooveMammalian CellMedical ResearchMethodsModernizationMycoplasmaMycoplasma hyorhinisOrganismOutputPerforationPhasePolymerasePrevalencePriceProtocols documentationRNARNA amplificationRNA-Directed DNA PolymeraseReactionReagentReportingReproducibilityResearchRibosomal RNASamplingScientistShipsSmall Business Innovation Research GrantSodium ChlorideSodium HydroxideSourceSpeedSpottingsSystemTechniquesTemperatureTestingTimeTubeUnited States National Institutes of HealthViolaamplification detectionaqueousassay developmentbasebiological researchcommercial applicationcomputerized data processingcostdesignexperimental studyinnovationluminescencenovelnovel strategiespathogenpathogenic bacteriapathogenic virusprototyperapid detectionrecombinaseremote locationresearch facilitytissue culturetriphenylmethane
项目摘要
SBIR 2020: 6823823214: The development of FastMyco™: A novel isothermal colorimetric assay for the rapid detection of mycoplasma contamination.
The development of FastMyco™: A novel isothermal colorimetric assay for the rapid detection
of mycoplasma contamination.
Project Summary/ Abstract
(Word Count: 350)
Mycoplasma (myco) contamination of mammalian cell lines are recognized as a major contributor to the lack of
reproducibility in modern biomedical research. Despite the availability of commercially marketed detection
systems, the unabated prevalence of myco in research attests to the shortcomings of these protocols. Indeed,
all currently marketed myco tests fail to address the actual needs of the consumer, the bench scientist.
For a myco detection product to be widely adopted it must: 1) require no additional equipment or infrastructure,
2) integrate seamlessly into a laboratory’s daily tissue culture routine with no additional labor burden on the
users, and 3) be sensitive, inexpensive, and yield results immediately. To this end, in Phase I of this proposal
we will optimize the FastMyco™ assay to test cell-culture media samples. FastMyco™ uses breakthrough
Recombinase Polymerase Amplification (RPA) coupled to an initial reverse transcriptase (RT) step (RT-RPA),
resulting in an all-in-one isothermal alternative to PCR myco detection. In FastMyco™, the RT-RPA reaction
will target the 16S rRNA of myco, with amplification conducted in the cell-culture incubator at 37°C for approx.
15- 20 minutes. The assay has a potential detection limit of less than 1 CFU/ ml and immediate colorimetric
output from an onboard detection system. The thermal stability of the system is a major technical and cost
advantage of FastMyco™ over comparable systems. This will be achieved using cutting-edge lyophilization
techniques that will create a multi-layered bead containing all necessary enzymes and reagents, freeze-dried
around a core of DNA-detection substrate. The bead dissolves at different rates releasing the detection
reagent only after target amplification has begun. In Phase II, we will continue to develop FastMyco™ for large
HTP research organizations but also expand the bead-based detection systems to other human and
agricultural pathogens. We will strive to integrate FastMyco™ into every laboratory’s routine cell-culture. The
assay would also give regulatory agencies such as NIH and FDA the leverage to require this more rigorous
testing protocols to help curb the spread of mycoplasma in research.
SBIR 2020:6823823214:FastMyco™的开发:一种用于快速检测支原体污染的新型等温比色法。
FastMyco™的开发:一种用于快速检测的新型等温比色法
支原体污染
项目总结/摘要
(Word数量:350)
哺乳动物细胞系的支原体(myco)污染被认为是缺乏
现代生物医学研究的可重复性。尽管商业化销售的检测方法
系统中,在研究中,myco的不减的流行证明了这些协议的缺点。的确,
所有目前市售的Myco测试都不能解决消费者、实验室科学家的实际需要。
对于广泛采用的myco检测产品,它必须:1)不需要额外的设备或基础设施,
2)无缝集成到实验室的日常组织培养程序中,
用户,以及3)敏感,廉价,并立即产生结果。为此,在本提案的第一阶段,
我们将优化FastMyco™测定以测试细胞培养基样品。FastMyco™采用突破性的
与初始逆转录酶(RT)步骤(RT-RPA)偶联的酶聚合酶扩增(RPA),
导致PCR真菌检测的一体化等温替代方案。在FastMyco™中,RT-RPA反应
将靶向myco的16 S rRNA,在37°C的细胞培养箱中进行扩增约10分钟。
15-20分钟该测定法具有小于1 CFU/ ml的潜在检测限,
来自机载检测系统的输出。系统的热稳定性是一个主要的技术和成本
FastMyco™相对于同类系统的优势。这将通过使用最先进的冻干技术来实现
技术,将创建一个多层珠含有所有必要的酶和试剂,冷冻干燥
围绕着DNA检测基质的核心珠子以不同的速率溶解,
只有在目标扩增开始后才使用试剂。在第二阶段,我们将继续开发FastMyco™,
HTP研究组织还将基于珠子的检测系统扩展到其他人类和
农业病原体我们将努力将FastMyco™整合到每个实验室的常规细胞培养中。的
检测也将给予NIH和FDA等监管机构更严格的要求,
测试协议,以帮助遏制支原体在研究中的传播。
项目成果
期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Skin Immuno-CometChip in 3D vs. 2D Cultures to Screen Topical Toxins and Skin-Specific Cytochrome Inducers.
皮肤免疫彗星芯片在 3D 与 2D 培养中筛选局部毒素和皮肤特异性细胞色素诱导剂。
- DOI:10.3390/genes14030630
- 发表时间:2023-03-02
- 期刊:
- 影响因子:3.5
- 作者:
- 通讯作者:
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{{ truncateString('DEAN ROSENTHAL', 18)}}的其他基金
Development of the UValidate platform for the profiling of topically applied chemical agents.
开发 UValidate 平台,用于分析局部应用的化学制剂。
- 批准号:
10484288 - 财政年份:2022
- 资助金额:
$ 27.43万 - 项目类别:
Development of the UValidate platform for the profiling of topically applied chemical agents.
开发 UValidate 平台,用于分析局部应用的化学制剂。
- 批准号:
10707098 - 财政年份:2022
- 资助金额:
$ 27.43万 - 项目类别:
Optimization of the UValidate platform to measure genotoxicity associated with current problematic UV chemical blockers
优化 UValidate 平台以测量与当前有问题的紫外线化学阻断剂相关的遗传毒性
- 批准号:
10338776 - 财政年份:2021
- 资助金额:
$ 27.43万 - 项目类别:
Id proteins in UV-mediated keratinocyte apoptosis
紫外线介导的角质形成细胞凋亡中的 Id 蛋白
- 批准号:
6884012 - 财政年份:2004
- 资助金额:
$ 27.43万 - 项目类别:
Id proteins in UV-mediated keratinocyte apoptosis
紫外线介导的角质形成细胞凋亡中的 Id 蛋白
- 批准号:
7031025 - 财政年份:2004
- 资助金额:
$ 27.43万 - 项目类别:
Id proteins in UV-mediated keratinocyte apoptosis
紫外线介导的角质形成细胞凋亡中的 Id 蛋白
- 批准号:
7356064 - 财政年份:2004
- 资助金额:
$ 27.43万 - 项目类别:
Id proteins in UV-mediated keratinocyte apoptosis
紫外线介导的角质形成细胞凋亡中的 Id 蛋白
- 批准号:
6729489 - 财政年份:2004
- 资助金额:
$ 27.43万 - 项目类别:
Id proteins in UV-mediated keratinocyte apoptosis
紫外线介导的角质形成细胞凋亡中的 Id 蛋白
- 批准号:
7216356 - 财政年份:2004
- 资助金额:
$ 27.43万 - 项目类别:
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