2022-04-12 カリフォルニア大学校アーバイン校(UCI)
・単純な微生物ループを用いて採取した唾液サンプルを、RT-PCRやLAMPなどの一般的で正確なDNA増幅技術を用いて、1本の採取管で直接分析し、ウイルスを検出できることを示した。
・サンプル採取、抽出、精製、更なる操作のいくつかのステップを省くことで、プロセスを大幅に合理化し、加速しました。” 彼らはまた、一般的な洗剤を使ってサンプルを変性させることで、それが非感染性になり、取り扱いが非常に簡単になることを発見しました。
<関連情報>
- https://news.uci.edu/2022/04/12/uci-team-develops-simplified-faster-testing-process-for-detecting-viral-rna/
- https://www.nature.com/articles/s41598-022-07871-4
ARS-CoV-2唾液検査用臨床サンプルのシングルチューブ採取と核酸分析 Single-tube collection and nucleic acid analysis of clinical samples for SARS-CoV-2 saliva testing
Kyle H. Cole,Alexis Bouin,Caila Ruiz,Bert L. Semler,Matthew A. Inlay &Andrej Lupták
Scientific Reports Published: 10 March 2022
DOI:https://doi.org/10.1038/s41598-022-07871-4
Abstract
The SARS-CoV-2 pandemic has brought to light the need for expedient diagnostic testing. Cost and availability of large-scale testing capacity has led to a lag in turnaround time and hindered contact tracing efforts, resulting in a further spread of SARS-CoV-2. To increase the speed and frequency of testing, we developed a cost-effective single-tube approach for collection, denaturation, and analysis of clinical samples. The approach utilizes 1 µL microbiological inoculation loops to collect saliva, sodium dodecyl sulfate (SDS) to inactivate and release viral genomic RNA, and a diagnostic reaction mix containing polysorbate 80 (Tween 80). In the same tube, the SDS-denatured clinical samples are introduced to the mixtures containing all components for nucleic acids detection and Tween 80 micelles to absorb the SDS and allow enzymatic reactions to proceed, obviating the need for further handling of the samples. The samples can be collected by the tested individuals, further decreasing the need for trained personnel to administer the test. We validated this single-tube sample-to-assay method with reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) and discovered little-to-no difference between Tween- and SDS-containing reaction mixtures, compared to control reactions. This approach reduces the logistical burden of traditional large-scale testing and provides a method of deployable point-of-care diagnostics to increase testing frequency.
