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使用氨甲环酸和紫外线A光灭活人血浆中的中东呼吸综合征冠状病毒

Inactivation of Middle East respiratory syndrome-coronavirus in human plasma using amotosalen and ultraviolet A light.

作者信息

Hindawi Salwa I, Hashem Anwar M, Damanhouri Ghazi A, El-Kafrawy Sherif A, Tolah Ahmed M, Hassan Ahmed M, Azhar Esam I

机构信息

Blood Transfusion Services, King Abdulaziz University Hospital, King Abdulaziz University, Jeddah, Saudi Arabia.

Department of Medicine, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia.

出版信息

Transfusion. 2018 Jan;58(1):52-59. doi: 10.1111/trf.14422. Epub 2017 Dec 14.

Abstract

BACKGROUND

Middle East respiratory syndrome-coronavirus (MERS-CoV) is a novel zoonotic pathogen. Although the potential for MERS-CoV transmission through blood transfusion is not clear, MERS-CoV was recognized as a pathogen of concern for the safety of the blood supply especially after its detection in whole blood, serum, and plasma of infected individuals. Here we investigated the efficacy of amotosalen and ultraviolet A light (UVA) to inactivate MERS-CoV in fresh-frozen plasma (FFP).

STUDY DESIGN AND METHODS

Pooled FFP units were spiked with a recent clinical MERS-CoV isolate. Infectious and genomic viral titers were determined in plasma before and after inactivation with amotosalen/UVA treatment by plaque assay and reverse transcription-quantitative polymerase chain reaction, respectively. In addition, residual replicating or live virus after inactivation was examined by passaging in the permissive Vero E6 cells.

RESULTS

The mean MERS-CoV infectious titer in pretreatment samples was 4.67 ± 0.25 log plaque-forming units (pfu)/mL, which was reduced to undetectable levels after inactivation with amotosalen/UVA demonstrating a mean log reduction of more than 4.67 ± 0.25 pfu/mL. Furthermore, inoculation of inactivated plasma on Vero E6 cells did not result in any cytopathic effect (CPE) even after 7 days of incubation and three consecutive passages, nor the detection of MERS RNA compared to pretreatment samples which showed complete CPE within 2 to 3 days postinoculation and log viral RNA titer ranging from 9.48 to 10.22 copies/mL in all three passages.

CONCLUSION

Our data show that amotosalen/UVA treatment is a potent and effective way to inactivate MERS-CoV infectious particles in FFP to undetectable levels and to minimize the risk of any possible transfusion-related MERS-CoV transmission.

摘要

背景

中东呼吸综合征冠状病毒(MERS-CoV)是一种新型人畜共患病原体。尽管MERS-CoV通过输血传播的可能性尚不清楚,但MERS-CoV被认为是血液供应安全方面值得关注的病原体,尤其是在其在感染个体的全血、血清和血浆中被检测到之后。在此,我们研究了补骨脂素和紫外线A光(UVA)对新鲜冷冻血浆(FFP)中MERS-CoV的灭活效果。

研究设计和方法

将近期临床分离的MERS-CoV毒株加入混合的FFP单位中。在用补骨脂素/UVA处理灭活前后,分别通过空斑试验和逆转录定量聚合酶链反应测定血浆中的感染性病毒滴度和基因组病毒滴度。此外,通过在允许生长的Vero E6细胞中传代来检测灭活后残留的复制性或活病毒。

结果

预处理样品中MERS-CoV的平均感染性滴度为4.67±0.25 log空斑形成单位(pfu)/mL,在用补骨脂素/UVA灭活后降至检测不到的水平,平均对数降低超过4.67±0.25 pfu/mL。此外,即使在孵育7天和连续传代三次后,将灭活血浆接种到Vero E6细胞上也未产生任何细胞病变效应(CPE),与预处理样品相比也未检测到MERS RNA,预处理样品在接种后2至3天内出现完全CPE,并且在所有三次传代中病毒RNA滴度对数范围为9.48至10.22拷贝/mL。

结论

我们的数据表明,补骨脂素/UVA处理是一种有效且有力的方法,可将FFP中的MERS-CoV感染性颗粒灭活至检测不到的水平,并将任何可能的输血相关MERS-CoV传播风险降至最低。

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