期刊
ANALYTICAL METHODS
卷 14, 期 47, 页码 5012-5018出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/d2ay01330d
关键词
-
资金
- National Natural Science Foundation of China
- Fundamental Research Funds for the Central Universities
- [21775012]
- [FRF-BR-20-03B]
- [FRF-TP-18-052A1]
This paper presents a novel visual reverse transcription-multiple inner primer loop-mediated isothermal amplification (RT-MIPLAMP) method based on RT-LAMP, which has higher sensitivity and shorter reaction time. The feasibility of visual RT-MIPLAMP for SARS-CoV-2 identification is validated.
Since the end of 2019, outbreaks of COVID-19 pandemics have continued in different areas worldwide, which exacerbates the need for rapid, sensitive and simple methods for diagnosis. Currently, COVID-19 diagnosis mainly relies on reverse transcription-polymerase chain reaction (RT-PCR), which requires sophisticated instruments. Reverse transcription-loop mediated isothermal amplification (RT-LAMP), due to its isothermal nature and high specificity, can be used as an alternative. In this paper, a novel visual reverse transcription-multiple inner primer loop-mediated isothermal amplification (RT-MIPLAMP) method is established based on RT-LAMP by adding a pair of inner primers. The RT-MIPLAMP method has a higher sensitivity and shorter reaction time compared with conventional RT-LAMP. By using RT-MIPLAMP, as low as 6 x 10(3) copies per mL in vitro transcribed (IVT) N gene can be detected within 55 min. Meanwhile, as low as 6 x 10(4) copies per mL IVT N gene is detectable with conventional RT-LAMP within 80 min. The feasibility of visual RT-MIPLAMP is also validated by detecting the N gene spiked into one healthy volunteer's saliva and the full-length RNA in pseudoviruses, indicating the great potential of visual RT-MIPLAMP for SARS-CoV-2 identification.
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