Digital PCR to quantify ChAdOx1 nCoV-19 copies in blood and tissues.

Digital PCR to quantify ChAdOx1 nCoV-19 copies in blood and tissues.
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DOI:
10.1016/j.omtm.2021.10.002
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发表时间:
2021-12-10
期刊:
Molecular therapy. Methods & clinical development
影响因子:
--
通讯作者:
Fehse B
Fehse B
中科院分区:
其他
文献类型:
--
作者:
Badbaran A;Mailer RK;Dahlke C;Woens J;Fathi A;Mellinghoff SC;Renné T;Addo MM;Riecken K;Fehse B

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使用基于腺病毒载体的AstraZeneca ChAdOx 1 nCov-19(Vaxzevria)疫苗接种是有效和安全的。然而,在极少数情况下,接种疫苗的个体发生危及生命的血栓性并发症,包括脑静脉窦和内脏静脉血栓形成。监测应用的载体在体内是一个重要的先决条件,以研究潜在的疫苗驱动的不良反应,现在被称为疫苗诱导的免疫血栓性血小板减少症(VITT)的分子机制。我们以前已经表明,数字PCR(dPCR)是一个很好的工具,定量转基因拷贝在体内。在这里,我们提出了一个高度敏感的dPCR原位定量ChAdOx 1 nCoV-19拷贝。使用该方法,我们定量了疫苗接种后24、72和168 h人血浆中的载体拷贝,以及注射后30 min实验疫苗接种模型中各种鼠组织中的载体拷贝。我们描述了一种高灵敏度定量检测ChAdOx 1 nCoV-19的方法,可能有助于阐明严重ChAdOx 1 nCoV-19疫苗并发症的机制。Badbaran等人引入用于量化ChAdOx 1 nCov-19疫苗载体拷贝数字PCR技术。基于其出色的特异性和灵敏度,新检测有助于在不同生物标本中监测ChAdOx 1 nCov-19,因此可能有助于阐明罕见但可能严重的疫苗引起的并发症的机制。
Vaccination with the adenoviral-vector-based AstraZeneca ChAdOx1 nCov-19 (Vaxzevria) vaccine is efficient and safe. However, in rare cases vaccinated individuals developed life-threatening thrombotic complications, including thrombosis in cerebral sinus and splanchnic veins. Monitoring of the applied vector in vivo represents an important precondition to study the molecular mechanisms underlying vaccine-driven adverse effects now referred to as vaccine-induced immune thrombotic thrombocytopenia (VITT). We previously have shown that digital PCR (dPCR) is an excellent tool to quantify transgene copies in vivo. Here, we present a highly sensitive dPCR for in situ quantification of ChAdOx1 nCoV-19 copies. Using this method, we quantified vector copies in human plasma 24, 72, and 168 h post vaccination and in a variety of murine tissues in an experimental vaccination model 30 min post injection. We describe a method for high-sensitivity quantitative detection of ChAdOx1 nCoV-19 with possible implications to elucidate the mechanisms of severe ChAdOx1 nCov-19 vaccine complications. Badbaran et al. introduce a digital PCR technique for quantification of ChAdOx1 nCov-19 vaccine vector copies. Based on its excellent specificity and sensitivity, the new assay facilitates ChAdOx1 nCov-19 monitoring in different biological specimens and might thus help to elucidate the mechanisms underlying the rare but potentially severe vaccine-caused complications.
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