A rapid and flexible microneutralization assay for serological assessment of influenza viruses.
A rapid and flexible microneutralization assay for serological assessment of influenza viruses.
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DOI：
10.1111/irv.13141
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发表时间：
2023
影响因子：
4.4
通讯作者：
Lauring, Adam S. S.
中科院分区：
文献类型：
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作者：
Rumfelt, Kalee E. E.;Fitzsimmons, William J. J.;Truscon, Rachel;Monto, Arnold S. S.;Martin, Emily T. T.;Lauring, Adam S. S.
流感疫苗接种或感染的血清学反应通常通过血凝素抑制（HAI）或微量中和（MN）来测量。这两种方法在可行性、标准化和推广到最近的菌株方面都有局限性。我们开发了一种荧光素酶MN （LMN）测定法，它结合了传统MN测定法的优点和HAI测定法的易用性。血清来自密歇根家庭队列的HIVE研究。利用反向遗传学技术产生重组流感病毒，表达测试菌株的血凝素和神经氨酸酶，所有其他病毒蛋白来自A/WSN/1933主干，以及NanoLuc报告基因。荧光素酶表达靶标的血清中和被量化为感染细胞的发光减少。测定了A/Hong Kong/4801/2014和A/Singapore/INFIMH‐16‐0019/2016的细胞适应型和卵适应型的中和效价，并与卵生长抗原的HAI效价进行了比较。2016年5月至2018年7月期间，从259名参与者中收集了333份血清。抽样的参与者年龄在7-68岁之间，其中80%接种了流感疫苗。A/Hong Kong/4801/2014和A/Singapore/INFIMH‐16‐0019/2016的HAI滴度与LMN滴度相关（ρ = 0.52, p≤0.01）。细胞株的LMN滴度低于卵株（A/Hong Kong/4801/2014平均log2倍变化=−2.66,p≤0.01，A/Singapore/INFIMH‐16‐0019/2016平均log2倍变化=−3.15,p≤0.01）。LMN试验在有限的样本量下是可行的，并且能够区分卵适应株和细胞衍生株之间的微小抗原差异。这些结果与常用的HAI相对应，证实了该试验在高通量研究保护和疫苗反应相关因素方面的实用性。
Serological responses from influenza vaccination or infection are typically measured by hemagglutinin inhibition (HAI) or microneutralization (MN). Both methods are limited in feasibility, standardization, and generalizability to recent strains. We developed a luciferase MN (LMN) assay that combines the advantages of the conventional MN assay with the ease of the HAI assay. Sera were obtained from the HIVE study, a Michigan household cohort. Reverse genetics was used to generate recombinant influenza viruses expressing the hemagglutinin and neuraminidase of test strains, all other viral proteins from an A/WSN/1933 backbone, and a NanoLuc reporter. Serum neutralization of luciferase‐expressing targets was quantified as a reduction in light emission from infected cells. Neutralization titers were measured for cell‐ and egg‐adapted versions of A/Hong Kong/4801/2014 and A/Singapore/INFIMH‐16‐0019/2016 and compared to HAI titers against egg‐grown antigens. Three hundred thirty‐three sera were collected from 259 participants between May 2016 and July 2018. Sampled participants were 7–68 years of age, and >80% were vaccinated against influenza. HAI and LMN titers were correlated for A/Hong Kong/4801/2014 (ρ = 0.52, p ≤ 0.01) and A/Singapore/INFIMH‐16‐0019/2016 (ρ = 0.79, p ≤ 0.01). LMN titers were lower for cell strains compared to egg strains (A/Hong Kong/4801/2014 mean log2 fold change = −2.66, p ≤ 0.01 and A/Singapore/INFIMH‐16‐0019/2016 mean log2 fold change = −3.15, p ≤ 0.01). The LMN assay was feasible using limited sample volumes and able to differentiate small antigenic differences between egg‐adapted and cell‐derived strains. The correspondence of these results with the commonly used HAI confirms the utility of this assay for high‐throughput studies of correlates of protection and vaccine response.
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影响因子：
5.5
作者：
Jackson ML;Phillips CH;Wellwood S;Kiniry E;Jackson LA;Martin ET;Monto AS;McLean HQ;Belongia EA;Gaglani M;Dunnigan K;Raiyani C;Murthy K;Flannery B;Chung JR
通讯作者：
Chung JR
DOI：
10.1093/cid/ciaa1884
发表时间：
2021-12-06
期刊：
Clinical infectious diseases : an official publication of the Infectious Diseases Society of America
影响因子：
--
作者：
Tenforde, Mark W;Kondor, Rebecca J Garten;Flannery, Brendan
通讯作者：
Flannery, Brendan
影响因子：
11.8
作者：
Chung, Jessie R.;Rolfes, Melissa A.;Reed, Carrie
通讯作者：
Reed, Carrie
影响因子：
9.4
作者：
OKUNO, Y;TANAKA, K;UEDA, S
通讯作者：
UEDA, S
影响因子：
4.4
作者：
Truelove S;Zhu H;Lessler J;Riley S;Read JM;Wang S;Kwok KO;Guan Y;Jiang CQ;Cummings DA
通讯作者：
Cummings DA