Sample preparation, including bacterial lysis, remains a hurdle in the realization of complete point-of-care tests for many pathogens. Here, we developed a sample preparation methodology for enzymatic lysis and sample heating for low-resource, point-of-care applications. We show an instrument-free chemical heater system for rapid lysis of a gram-positive bacterium (Staphylococcus aureus) and an RNA virus (human respiratory syncytial virus) using a dried lysis enzyme mixture (achromopeptidase) for S. aureus. After a lysis step (<1 minute), lysis enzymes are heat deactivated (<5 minutes) using a simple disposable chemical heater. We demonstrated that both DNA and RNA in the heat-treated sample could be directly amplified without purification, even in the presence of a clinically-obtained human nasal sample. This simple approach to dry enzyme storage and sample heating is adaptable to many applications where samples need to be lysed, including use in low-resource laboratories and in single-use or cartridge-based point-of-care diagnostic devices.
A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms.
用于裂解和提取微生物DNA和RNA的化学加热器和干酶制剂
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作者:Buser J R, Zhang X, Byrnes S A, Ladd P D, Heiniger E K, Wheeler M D, Bishop J D, Englund J A, Lutz B, Weigl B H, Yager P
| 期刊: | Analytical Methods | 影响因子: | 2.600 |
| 时间: | 2016 | 起止号: | 2016 Apr 14; 8(14):2880-2886 |
| doi: | 10.1039/c6ay00107f | 研究方向: | 微生物学 |
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