Improved quantification of a commercial enzyme-linked immunosorbent assay kit for measuring anti-MDA5 antibody
Improved quantification of a commercial enzyme-linked immunosorbent assay kit for measuring anti-MDA5 antibody
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DOI：
10.1080/14397595.2018.1452179
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发表时间：
2019-01-02
影响因子：
2.2
通讯作者：
Kuwana, Masataka
中科院分区：
文献类型：
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作者：
Gono, Takahisa;Okazaki, Yuka;Kuwana, Masataka
目的：比较两种酶联免疫吸附试验（ELISA）系统测定抗MDA 5抗体滴度的定量性能：内部ELISA和商业MESACUP（TM）抗MDA 5试验。研究方法：使用内部ELISA和MESACUPTM抗-MDA 5测试并行测量来自70名皮肌炎患者的血清中的抗-MDA 5抗体滴度。对于商业ELISA试剂盒，根据制造商的方案使用1：101稀释的血清样品，但也检查血清的系列稀释液以鉴定用于定量的最佳血清稀释度。结果如下：通过内部和商业ELISA测量的抗-MDA 5抗体滴度彼此正相关（r = 0.53，p = .0001），但通过商业ELISA测量的抗体滴度对药物治疗后的变化不太敏感，46份抗-MDA 5阳性血清中有37份（80%）的抗体效价超过了厂家规定的定量范围（≥ 150指数）。使用稀释的血清样品进行的实验表明，将血清稀释1：5050改善了MESACUPTM抗MDA 5测试的定量性能，包括与内部ELISA结果更好的相关性和对变化的敏感性增加。结论：通过改进ELISA试剂盒的检测方法，提高了该试剂盒检测抗MDA 5抗体效价的能力。
Objectives: To compare the quantitative performance for measuring anti-MDA5 antibody titer of two enzyme-linked immunosorbent assay (ELISA) systems: an in-house ELISA and the commercial MESACUP(TM) anti-MDA5 test. Methods: Anti-MDA5 antibody titer was measured in sera from 70 patients with dermatomyositis using an in-house ELISA and the MESACUP(TM) anti-MDA5 test side-by-side. For the commercial ELISA kit, serum samples diluted 1:101 were used according to the manufacturer's protocol, but serial dilutions of sera were also examined to identify the optimal serum dilution for quantification. Results: The anti-MDA5 antibody titers measured by the in-house and commercial ELISAs were positively correlated with each other (r = 0.53, p = .0001), but the antibody titer measured by the commercial ELISA was less sensitive to change after medical treatment, and 37 (80%) of 46 anti-MDA5-positive sera had antibody titer exceeding the quantification range specified by the manufacturer (>= 150 index). Experiments using diluted serum samples revealed that diluting the sera 1:5050 improved the quantitative performance of the MESACUP(TM) anti-MDA5 test, including a better correlation with the in-house ELISA results and an increased sensitivity to change. Conclusion: We improved the ability of the commercial ELISA kit to quantify anti-MDA5 antibody titer by altering its protocol.