Stabilization of Tuberculosis Reporter Enzyme Fluorescence (REFtb) Diagnostic Reagents for Use at the Point of Care.
Stabilization of Tuberculosis Reporter Enzyme Fluorescence (REFtb) Diagnostic Reagents for Use at the Point of Care.
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DOI：
10.3390/diagnostics12071745
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发表时间：
2022-07-19
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影响因子：
--
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结核病是全世界人类最常见的死亡原因之一。结核病仍然是一个公共卫生威胁的主要原因之一是，诊断可能需要数周到数月的时间，通常不是很敏感，并且在许多偏远环境中无法完成。快速、敏感且廉价的护理点（POC）诊断将对根除结核病的工作产生重大影响。结核病诊断系统REFtb基于使用指定为CDG-3的定制荧光底物特异性检测结核分枝杆菌中组成型表达的β-内酰胺酶（BlaC）。REFtb有潜力作为结核病的诊断试剂，可能非常便宜（<2.00美元/测试），在POC中使用，可以在10分钟内提供明确的诊断。然而，REFtb的试剂目前是液体形式，使它们更容易降解，难以运输。我们通过在各种条件下冻干对REFtb性能的影响，评价了REFtb试剂稳定性的改善。我们发现，冷冻干燥REFtb组分可产生易于重构的粉末，其表现出与液体系统相似的性能，并且乳糖是用于最终POC REFtb诊断系统的最有前途的赋形剂之一。这些研究为生产稳定的POC REFtb系统提供了基础，该系统可以在全球范围内轻松分销，对制冷的要求最低或不要求制冷。
Tuberculosis is one of the most frequent causes of death in humans worldwide. One of the primary reasons tuberculosis remains a public health threat is that diagnosis can take weeks to months, is often not very sensitive and cannot be accomplished in many remote environments. A rapid, sensitive and inexpensive point-of-care (POC) diagnostic would have a major impact on tuberculosis eradication efforts. The tuberculosis diagnostic system REFtb is based on specific detection of the constitutively expressed β-lactamase (BlaC) in Mycobacterium tuberculosis using a custom fluorogenic substrate designated as CDG-3. REFtb has potential as a diagnostic for tuberculosis that could be very inexpensive (<USD 2.00/test), used at the POC and could provide definitive diagnosis within 10 min. However, the reagents for REFtb are currently in liquid form, making them more susceptible to degradation and difficult to transport. We evaluated the improvement in the stability of REFtb reagents by lyophilization under a variety of conditions through their effects on the performance of REFtb. We found that lyophilization of REFtb components produces an easily reconstituted powder that displays similar performance to the liquid system and that lactose represents one of the most promising excipients for use in a final POC REFtb diagnostic system. These studies provide the foundation for the production of a stable POC REFtb system that could be easily distributed worldwide with minimal or no requirement for refrigeration.
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DOI：
10.1073/pnas.1000643107
发表时间：
2010-07-06
影响因子：
11.1
作者：
Kong, Ying;Yao, Hequan;Cirillo, Jeffrey D.
通讯作者：
Cirillo, Jeffrey D.
DOI：
10.1016/j.tube.2016.09.009
发表时间：
2016-12
期刊：
Tuberculosis (Edinburgh, Scotland)
影响因子：
--
作者：
Sule P;Tilvawala R;Behinaein P;Walkup GK;Cirillo JD
通讯作者：
Cirillo JD
影响因子：
10
作者：
HEILPERN, S;REBUCK, AS
通讯作者：
REBUCK, AS
影响因子：
11.8
作者：
Getahun, Haileyesus;Gunneberg, Christian;Nunn, Paul
通讯作者：
Nunn, Paul
DOI：
10.1093/infdis/jiw298
发表时间：
2017-02-01
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
Yang HJ;Kong Y;Cheng Y;Janagama H;Hassounah H;Xie H;Rao J;Cirillo JD
通讯作者：
Cirillo JD