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在PrimeStore分子转运介质中成功灭活高致病性病原体

Successful Inactivation of High-Consequence Pathogens in PrimeStore Molecular Transport Media.

作者信息

Spruill-Harrell Briana, Kocher Gregory, Boda Maurice, Akers Kristen, Freeburger Denise, Murphy Nicole, Kuhn Jens H, Fischer Gerald, Maljkovic Berry Irina, Chandrasekaran Prabha, Torrison Jerry

机构信息

Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, B-8200 Research Plaza, Fort Detrick, Frederick, MD 21702, USA.

Longhorn Vaccines and Diagnostics, LLC, Bethesda, MD 20814, USA.

出版信息

Viruses. 2025 Apr 29;17(5):639. doi: 10.3390/v17050639.

Abstract

Handling cultured isolates and clinical, environmental, or wildlife surveillance samples containing Risk Group 3 and 4 pathogens presents considerable biosafety challenges in minimizing human exposure during processing and transport. Safe handling typically requires high- or maximum-containment facilities, demanding substantial logistical planning and resources. We evaluated PrimeStore Molecular Transport Medium (PS-MTM), a guanidine-based solution created to kill pathogens and preserve nucleic acids at ambient temperatures, for inactivating Crimean-Congo hemorrhagic fever, eastern equine encephalitis, Ebola, Hendra, Japanese encephalitis, Lassa, Marburg, Nipah, Rift Valley fever, and West Nile viruses. To mimic diagnostic conditions, human whole blood spiked with any of these viruses was incubated with PS-MTM for 20-, 30-, or 60-min. Samples with titers up to 10 PFU/mL exposed to PS-MTM at all time points resulted in complete loss of infectivity judged by plaque assays. A 30-min incubation provided a 50% safety margin over the minimum inactivation time and was used for quantification with the tissue culture infectious dose (TCID) assay, enabling evaluation of PS-MTM's activity for viruses that do or do not produce well-defined plaques. Results confirmed that PS-MTM inactivated all tested viruses at titers up to 10 TCID/mL, underscoring its reliability for enhancing biosafety in diagnostics, outbreak management, and surveillance.

摘要

处理含有风险等级3和4病原体的培养分离株以及临床、环境或野生动物监测样本,在处理和运输过程中尽量减少人员接触方面带来了相当大的生物安全挑战。安全处理通常需要高防护或最高防护设施,这需要大量的后勤规划和资源。我们评估了PrimeStore分子转运培养基(PS-MTM),这是一种基于胍的溶液,旨在在常温下杀灭病原体并保存核酸,用于灭活克里米亚-刚果出血热病毒、东部马脑炎病毒、埃博拉病毒、亨德拉病毒、日本脑炎病毒、拉沙病毒、马尔堡病毒、尼帕病毒、裂谷热病毒和西尼罗河病毒。为模拟诊断条件,将接种了上述任何一种病毒的人全血与PS-MTM孵育20分钟、30分钟或60分钟。在所有时间点,滴度高达10 PFU/mL的样本经PS-MTM处理后,通过空斑试验判断其感染性完全丧失。30分钟的孵育时间比最短灭活时间有50%的安全余量,并用于组织培养感染剂量(TCID)试验进行定量分析,从而能够评估PS-MTM对产生或不产生清晰空斑的病毒的活性。结果证实,PS-MTM能灭活滴度高达10 TCID/mL的所有测试病毒,突出了其在诊断、疫情管理和监测中增强生物安全方面的可靠性。

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