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使用防腐剂和抗生素以提高脊髓灰质炎病毒在带正电滤器上的存活时间

Use of Preservative Agents and Antibiotics for Increased Poliovirus Survival on Positively Charged Filters.

作者信息

Fagnant Christine Susan, Kossik Alexandra Lynn, Zhou Nicolette Angela, Sánchez-Gonzalez Liliana, Falman Jill Christin, Keim Erika Karen, Linden Yarrow, Scheibe Alana, Barnes Kilala Sayisha, Beck Nicola Koren, Boyle David S, Meschke John Scott

机构信息

Department of Environmental & Occupational Health Sciences, University of Washington, 4225 Roosevelt Way NE, Suite 100, Seattle, WA, 98105, USA.

PATH, 2201 Westlake Avenue, Suite 200, Seattle, WA, 98121, USA.

出版信息

Food Environ Virol. 2017 Dec;9(4):383-394. doi: 10.1007/s12560-017-9306-4. Epub 2017 Jun 14.

Abstract

Environmental surveillance of poliovirus (PV) and other non-enveloped viruses can help identify silent circulation and is necessary to certify eradication. The bag-mediated filtration system is an efficient method to filter large volumes of environmental waters at field sites for monitoring the presence of viruses. As filters may require long transit times to off-site laboratories for processing, viral inactivation or overgrowth of bacteria and fungi can interfere with virus detection and quantification (Miki and Jacquet in Aquatic Microb Ecol 51(2):195-208, 2008). To evaluate virus survival over time on ViroCap filters, the filters were seeded with PV type 1 (PV1) and/or MS2 and then dosed with preservatives or antibiotics prior to storage and elution. These filters were stored at various temperatures and time periods, and then eluted for PV1 and MS2 recovery quantification. Filters dosed with the preservative combination of 2% sodium benzoate and 0.2% calcium propionate had increased virus survival over time when stored at 25 °C, compared to samples stored at 25 °C with no preservatives. While elution within 24 h of filtration is recommended, if storage or shipping is required then this preservative mixture can help preserve sample integrity. Addition of an antibiotic cocktail containing cephapirin, gentamicin, and Proclin 300 increased recovery after storage at 4 and 25 °C, when compared to storage with no antibiotics. The antibiotic cocktail can aid sample preservation if access to appropriate antibiotics storage is available and sample cold chain is unreliable. This study demonstrated that the use of preservatives or antibiotics is a simple, cost-effective method to improve virus detection from ViroCap cartridge filters over time.

摘要

脊髓灰质炎病毒(PV)及其他无包膜病毒的环境监测有助于识别隐匿传播,对于认证根除工作至关重要。袋式介导过滤系统是一种在现场过滤大量环境水样以监测病毒存在的有效方法。由于过滤器可能需要较长时间运往异地实验室进行处理,病毒失活或细菌和真菌的过度生长会干扰病毒检测和定量(Miki和Jacquet,《水生微生物生态学》51(2):195 - 208,2008年)。为评估病毒在ViroCap过滤器上随时间的存活情况,在过滤器上接种1型脊髓灰质炎病毒(PV1)和/或MS2,然后在储存和洗脱前加入防腐剂或抗生素。这些过滤器在不同温度和时间段储存,然后洗脱以对PV1和MS2进行回收定量。与在25°C下未添加防腐剂储存的样品相比,添加2%苯甲酸钠和0.2%丙酸钙的防腐剂组合的过滤器在25°C储存时,病毒随时间的存活情况有所改善。虽然建议在过滤后24小时内进行洗脱,但如果需要储存或运输,这种防腐剂混合物有助于保持样品完整性。与未添加抗生素储存相比,添加含有头孢匹林、庆大霉素和Proclin 300的抗生素鸡尾酒可提高在4°C和25°C储存后的回收率。如果能够获得适当的抗生素储存条件且样品冷链不可靠,抗生素鸡尾酒有助于样品保存。本研究表明,使用防腐剂或抗生素是一种简单、经济有效的方法,可随着时间的推移提高从ViroCap滤芯过滤器中检测病毒的能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f911/5668339/ee9803b49b69/12560_2017_9306_Fig1_HTML.jpg

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