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Tatsuya Mikami,Yuta Saeki,Sayaka Hirai,Mayuko Shimokawa,Yukiko Umeyama,Yusaku Kuroda,Hiroaki Kodama 한국식물생명공학회 2018 Plant biotechnology reports Vol.12 No.6
RNA silencing is a sequence-specific form of epigenetic regulation that targets invasive nucleic acids. RNA-dependent RNA polymerase6 (RDR6) converts target RNA molecules, such as transgene transcripts, into double-stranded RNAs (dsRNAs) during posttranscriptional gene silencing (PTGS). Then, these dsRNAs are processed into small RNAs that guide sequencespecific RNA degradation. T-DNA-derived small RNAs are generated during the transfer of T-DNA from Agrobacterium to plant cells and compromise the function of the genes in the T-DNA. In the present study, we produced selection-markerfree transgenic tobacco plants using the MAT vector system, and expression of the tobacco RDR6 gene (NtRDR6) was suppressed using inverted-repeat-induced PTGS. Reduced expression of the NtRDR6 gene improved the transient expression of the transgene in the agroinfiltrated leaves and enhanced the production of hairy roots after infection with Agrobacterium containing a root-inducing T-DNA. The expression level of the sense transgene was determined in individual hairy roots, and knockdown of the NtRDR6 gene did not affect the distribution of the expression levels in individual transformants. These results indicate that NtRDR6 partially inhibited T-DNA function during T-DNA transfer but did not affect the expression of the transgene in stable transformants, except in transformants showing sense-transgene-induced PTGS.
Combination of a SARS-CoV-2 IgG Assay and RT-PCR for Improved COVID-19 Diagnosis
Aoki Kotaro,Takai Kunitomo,Nagasawa Tatsuya,Kashiwagi Katsuhito,Mori Nobuaki,Matsubayashi Keiji,Satake Masahiro,Tanaka Ippei,Kodama Nanae,Shimodaira Takahiro,Ishii Yoshikazu,Miyazaki Taito,Ishii Toshi 대한진단검사의학회 2021 Annals of Laboratory Medicine Vol.41 No.6
Background: Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is generally diagnosed by reverse transcription (RT)-PCR or serological assays. The SARS-CoV-2 viral load decreases a few days after symptom onset. Thus, the RT-PCR sensitivity peaks at three days after symptom onset (approximately 80%). We evaluated the performance of the ARCHITECT® SARS-CoV-2 IgG assay (henceforth termed IgG assay; Abbott Laboratories, Lake County, IL, USA), and the combination of RT-PCR and the IgG assay for COVID-19 diagnosis. Methods: In this retrospective study, 206 samples from 70 COVID-19 cases at two hospitals in Tokyo that were positive using RT-PCR were used to analyze the diagnostic sensitivity. RT-PCR-negative (N=166), COVID-19-unrelated (N=418), and Japanese Red Cross Society (N=100) samples were used to evaluate specificity. Results: Sensitivity increased daily after symptom onset and exceeded 84.4% after 10 days. Specificity ranged from 98.2% to 100% for samples from the three case groups. Seroconversion was confirmed from 9 to 20 days after symptom onset in 18 out of 32 COVID-19 cases with multiple samples and from another case with a positive result in the IgG assay for the first available sample. Conclusions: The combination of RT-PCR and IgG assay improves the robustness of laboratory diagnostics by compensating for the limitations of each method.