{"created":"2025-01-22T00:06:05.919644+00:00","id":2001685,"links":{},"metadata":{"_buckets":{"deposit":"65945f20-b60c-4503-90c9-843eb4f1e681"},"_deposit":{"created_by":10,"id":"2001685","owners":[10],"pid":{"revision_id":0,"type":"depid","value":"2001685"},"status":"published"},"_oai":{"id":"oai:repository.lib.tottori-u.ac.jp:02001685","sets":["1:9","2:12","23:34:1710471377862:1737507731107"]},"author_link":[],"item_10001_biblio_info_7":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2024-11-27","bibliographicIssueDateType":"Issued"},"bibliographicIssueNumber":"4","bibliographicNumberOfPages":"10","bibliographicPageEnd":"302","bibliographicPageStart":"293","bibliographicVolumeNumber":"67","bibliographic_titles":[{"bibliographic_title":"Yonago Acta Medica","bibliographic_titleLang":"en"}]}]},"item_10001_description_5":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Background Recently developed rapid real-time reverse transcription PCR (RT-PCR) systems adopting microfluidic thermal cycling technology are ideal forpoint-of-care (POC) testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Because the RNA extraction step before real-time RT-PCR is rate-limiting, a direct RNA extraction method (direct method) that adopts chemical viral lysis and eliminates RNA purification steps is preferable for rapid real-time RT-PCR. In the direct method, selecting the transport medium is essential because it may be introduced into subsequent real-time RT-PCR steps, but might inhibit PCR. However, the influence of transport medium on the combination of the direct method and rapid realtime RT-PCR has been yet unstudied. In the present study, we examined the influence of various transport mediums when combining the direct method and rapid real-time RT-PCR of GeneSoC® (GeneSoC® RT-PCR), the recently developed compact PCR system that adapts novel microfluidic thermal cycling technology. Methods To explore the influence of the transport medium on the GeneSoC® RT-PCR, the concordance of the RNA extraction and direct method was evaluated in the clinical samples collected in viral transport medium (VTM) or eSwab®. The sensitivity of GeneSoC® RTPCR combined with the direct method was assessed using spiked samples in generic (H2O and PBS) or commercially available transport media (VTM and eSwab®). Analytical sensitivity was examined using clinical specimens collected from the VTM and eSwab®. The inhibitory effect of PCR inhibitors on clinical specimens was assessed using clinical samples diluted 1,000 times. Results While only 1 copy/reaction of RNA was detected in H2O and eSwab® of the spiked samples, a minimum of 5 copies/reaction was detected in PBS (-) and VTM. Among the clinical specimens tested using the direct method, the detection of viral RNA was unstable in the samples containing less than 100 copies/reaction viral RNA in VTM, whereas less than 10 copies/reaction viral RNA were detected in eSwab®. The positive, negative, and overall concordance between the RNA extraction and the direct method was 84%, 100%, and 85%, respectively, in eSwab® samples, whereas the values were 35%, 100%, and 38%, respectively, in VTM samples. When the clinical samples were diluted 1,000 times, GeneSoC® RT-PCR could detect as low as 1.15 copies/reaction RNA using direct method, and the sensitivity was comparable to that of RNA extraction. Conclusion The combination of the direct method and microfluidic rapid PCR machine GeneSoC® has a high sensitivity for detecting SARS-CoV-2 RNA in clinical samples with eSwab® transport medium.","subitem_description_language":"en","subitem_description_type":"Other"}]},"item_10001_publisher_8":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"Tottori University Medical Press","subitem_publisher_language":"en"}]},"item_10001_relation_16":{"attribute_name":"情報源","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_language":"en","subitem_relation_name_text":"Yonago Acta Medica. 2024, 67(4), 293-302."}]}]},"item_10001_relation_17":{"attribute_name":"関連サイト","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://www.lib.tottori-u.ac.jp/yam/yam/yam67-4/67-4contents.html"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://www.lib.tottori-u.ac.jp/yam/yam/yam67-4/67-4contents.html","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://doi.org/10.33160/yam.2024.11.003"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://doi.org/10.33160/yam.2024.11.003","subitem_relation_type_select":"DOI"}}]},"item_10001_rights_15":{"attribute_name":"権利","attribute_value_mlt":[{"subitem_rights":"(C)2024 Tottori University Medical 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Rapid Microfluidic PCR System"],"weko_creator_id":"10","weko_shared_id":-1},"updated":"2025-02-13T02:21:32.812250+00:00"}