The coronavirus disease 2019 ) pandemic now has >2,000,000 confirmed cases worldwide. COVID-19 is currently diagnosed using quantitative RT-PCR methods, but the capacity of quantitative RT-PCR methods is limited by their requirement of high-level facilities and instruments. We developed and evaluated reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays to detect genomic RNA of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative virus of COVID-19. RT-LAMP assays reported in this study can detect as low as 100 copies of SARS-CoV-2 RNA. Cross-reactivity of RT-LAMP assays to other human coronaviruses was not observed. A colorimetric detection method was adapted for this RT-LAMP assay to enable higher throughput.
The Hippo pathway regulates the self-renewal and differentiation of various adult stem cells, but its role in cell fate determination and differentiation during liver development remains unclear. Here we report that the Hippo pathway controls liver cell lineage specification and proliferation separately from Notch signalling, using mice and primary hepatoblasts with liver-specific knockout of Lats1 and Lats2 kinase, the direct upstream regulators of YAP and TAZ. During and after liver development, the activation of YAP/TAZ induced by loss of Lats1/2 forces hepatoblasts or hepatocytes to commit to the biliary epithelial cell (BEC) lineage. It increases BEC and fibroblast proliferation by up-regulating TGFβ signalling, but suppresses hepatoblast to hepatocyte differentiation by repressing Hnf4α expression. Notably, oncogenic YAP/TAZ activation in hepatocytes induces massive p53-dependent cell senescence/death. Together, our results reveal that YAP/TAZ activity levels govern liver cell differentiation and proliferation in a context-dependent manner.
Coronavirus disease 2019 (COVID-19) is a newly emerging human
infectious disease caused by Severe Acute Respiratory Syndrome
Coronavirus 2 (SARS-CoV-2, also previously known as 2019-nCoV).
Within 8 months of the outbreak, more than 10,000,000 cases of
COVID-19 have been confirmed worldwide. Since human-to-human
transmission occurs easily and the rate of human infection is
rapidly increasing, sensitive and early diagnosis is essential
to prevent a global outbreak. Recently, the World Health
Organization (WHO) announced various primer–probe sets
for SARS-CoV-2 developed at different institutions: China Center
for Disease Control and Prevention (China CDC, China),
Charité (Germany), The University of Hong Kong (HKU, Hong
Kong), National Institute of Infectious Diseases in Japan (Japan
NIID, Japan), National Institute of Health in Thailand (Thailand
NIH, Thailand), and US CDC (USA). In this study, we compared the
ability to detect SARS-CoV-2 RNA among seven primer–probe
sets for the N gene and three primer–probe sets for the
Orf1 gene. The results revealed that
“NIID_2019-nCOV_N” from the Japan NIID and
“ORF1ab” from China CDC represent a recommendable
performance of RT-qPCR analysis for SARS-CoV-2 molecular
diagnostics without nonspecific amplification and
cross-reactivity for hCoV-229E, hCoV-OC43, and MERS-CoV RNA.
Therefore, the appropriate combination of NIID_2019-nCOV_N
(Japan NIID) and ORF1ab (China CDC) sets should be selected for
sensitive and reliable SARS-CoV-2 molecular diagnostics.
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