期刊
ANALYTICAL METHODS
卷 10, 期 45, 页码 5358-5363出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c8ay01475b
关键词
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资金
- U.S. Department of Health and Human Services, National Institutes of Health, National Institute of Nursing Research, Division of Intramural Research [1Z1ANR000018-01-7]
- National Center For Advancing Translational Sciences [R43TR001912]
- U.S. Department of Health and Human Services, National Institutes of Health, Intramural Research Training Awards
Diarrheal diseases claim the lives of 1300 children daily, mostly in the developing world. We have developed a simple lateral flow assay capable of detecting E. coli and EPEC DNA and RNA rapidly (<15 minutes) at the point-of-need, directly from stool without nucleic acid extraction or molecular amplification. The limit of detection of the method is 1 nM using synthetic DNA target substrates spiked into stool. However, due to the endogenous amplification of the 23S rRNA targets, we were able to detect the endogenous EPEC in pea-sized (5 mg) stool without labor-intensive and time-consuming nucleic acid purification or target amplification using enzymes. The significance of this method is that it is rapid (<15 minutes) and simple (without nucleic acid purification or molecular amplification) and does not require instrumentation, or access to a laboratory, cold chain or electric power. Thus, it is well-suited for point-of-need use in remote and/or resource-limited settings in the developing world where the mortality due to diarrheal diseases is especially high. The rapid testing of stool pathogens in real time at the point-of-need will decrease the loss of patients to follow-up, and enable patients to be treated earlier with the appropriate therapeutics in both the developed and developing world settings.
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