- 期刊
以SARS-CoV-2偽病毒株評估CMP^(TM)去活化型病毒運送培養基（I-VTM）滅活病毒與保存RNA之效能
Using SARS-CoV-2 Pseudovirus to Evaluate CMP^(TM) Inactivation Viral Transport Medium (I-VTM) for Virus Inactivation and Viral RNA Preservation
摘要
新型冠狀病毒（SARS-CoV-2）具有高傳染性，為降低檢驗人員操作潛在病毒檢體時的風險，有必要在不影響分子檢驗結果的情況下，於運送過程中對其進行滅活。本研究欲評估啟新生物科技股份有限公司開發的CMP^(TM)去活化型病毒運送培養基（CMP^(TM) Inactivation Viral Transport Medium, I-VTM）滅活SARS-CoV-2及保存RNA之效能。出於安全考量，吾等使用兩株複製缺陷的SARS-CoV-2偽病毒（pseudovirus）（購自中央研究院，台灣），套膜上分別表現有新冠病毒武漢株（Wuhan strain）和德耳塔株（Delta strain）的棘蛋白（spike protein），並且內含螢火蟲冷光素酶基因（firefly luciferase gene），可用以評估病毒感染力。測試時，分別將此兩偽病毒加入I-VTM中，於4℃和25℃環境下靜置5、30與120分鐘，一部分處理後的偽病毒與293T-ACE2細胞混合，以冷光素酶活性評估兩病毒株的感染力；另一部分經處理的偽病毒則直接進行RNA萃取，以反轉錄－即時聚合酶反應（quantitative reverse transcription PCR，qRT-PCR）檢測firefly luciferase gene，確認I-VTM對RNA的保存效能。結果指出，I-VTM能在4℃和25℃下，於5分鐘高效滅活此兩株偽病毒，並在長達2小時的室溫靜置後，仍能保留病毒RNA的完整性可用於qRT-PCR分子檢測。綜合而言，與傳統需在2-8℃運送病毒檢體相比，I-VTM不僅能有效減低檢測人員感染風險，更能在室溫下保存病毒核酸的完整性，展現此產品在臨床病毒檢測中的潛在應用價值。
並列摘要
As SARS-CoV-2 is highly contagious, it is necessary to inactivate the specimen during transportation without interfering the molecular detection result. In this study, we evaluated the effectiveness of the CMP^(TM) Inactivation Viral Transport Medium (I-VTM), developed by Creative Life Science Co, Ltd., in the inactivation of SARS-CoV-2 and preservation of viral RNA. For safety reasons, two strains of replication defective SARS-CoV-2 pseudoviruses were used during experiment (purchased from Academia Sinica, Taiwan). These pseudoviruses express the same spike protein of SARS-CoV-2 Wuhan or Delta strain, and containing the firefly luciferase gene for assessing viral infectivity. Two pseudoviruses placed in I-VTM were stored at 4℃ or 25℃ for 5, 30, 120 minutes. A part of sample were mixed with 293T-ACE2 cells and evaluate their infectivity based on luciferase activity. Others went through RNA extraction and quantitative reverse transcription PCR (qRT-PCR) to detect firefly luciferase gene expression as an assessment for RNA preservation. Results showed that I-VTM was able to inactivate both viruses within 5 minute at 4℃ or 25℃ and retained the integrity of viral RNA for qRT-PCR for 2 hour in 25℃. In conclusion, compared with the traditional transportation of viral specimens at 2-8℃, specimens collected with I-VTM can not only reduce the risk of infection during testing, but also preserve the integrity of viral RNA at room temperature without affecting the effectiveness of viral detection, demonstrating the potential of this product applicated in clinical virus detection.