Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, USA.
Lab Chip. 2019 Mar 27;19(7):1217-1225. doi: 10.1039/c8lc01404c.
We describe a Lab-on-a-Film disposable that detects multidrug-resistant tuberculosis (MDR-TB) from sputum extracts. The Lab-on-a-Film disposable consists of 203 gel elements that include DNA sequences (probes) for 37 mutations, deletions, or insertion elements across 5 genes (including an internal control). These gel elements are printed on a flexible film, which costs approximately 500 times less than microarray glass. The film with printed gel elements is then laminated to additional rollable materials (films) to form a microfluidic flow cell. We combined multiplex amplification and hybridization steps in a single microfluidic chamber, without buffer exchanges or other manipulations up to and throughout hybridization. This flow cell also incorporates post hybridization wash steps while retaining an entirely closed-amplicon system, thus minimizing the potential for sample or amplicon cross-contamination. We report analytical sensitivity of 32 cfu mL-1 across all MDR-TB markers and detection of MDR-TB positive clinical specimens using an automated TruTip workstation for extraction and the Lab-on-a-Film disposable for amplification and detection of the extracts.
我们描述了一种基于薄膜的一次性设备，可从痰液提取物中检测耐多药结核病（MDR-TB）。基于薄膜的一次性设备由 203 个凝胶元件组成，这些凝胶元件包括 5 个基因（包括内部对照）中的 37 个突变、缺失或插入元件的 DNA 序列（探针）。这些凝胶元件被打印在柔性薄膜上，其成本大约比微阵列玻璃低 500 倍。带有打印凝胶元件的薄膜然后层压到其他可滚动的材料（薄膜）上，以形成微流控流动池。我们在单个微流控室中组合了多重扩增和杂交步骤，无需进行缓冲液交换或其他操作，直到并贯穿整个杂交过程。该流动池还在保留完全封闭的扩增子系统的同时包含杂交后清洗步骤，从而最大程度地减少样品或扩增子交叉污染的可能性。我们报告了在所有 MDR-TB 标志物中具有 32 cfu mL-1 的分析灵敏度，并且使用自动化 TruTip 工作站进行提取以及基于薄膜的一次性设备进行扩增和检测提取物来检测 MDR-TB 阳性临床标本。
