Immunofluorescence assay for detection of human metapneumovirus-specific antibodies by use of baculovirus-expressed fusion protein
Immunofluorescence assay for detection of human metapneumovirus-specific antibodies by use of baculovirus-expressed fusion protein
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DOI：
10.1128/cdli.12.1.202-205.2005
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发表时间：
2005-01-01
期刊：
影响因子：
--
通讯作者：
Kikuta, H
中科院分区：
文献类型：
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作者：
Ishiguro, N;Ebihara, T;Kikuta, H
人偏肺病毒（Human metapneumovirus，hMPV）是近年来发现的一种急性呼吸道感染的病原体。hMPV融合（F）蛋白已被证明是介导有效中和和保护hMPV感染的主要抗原决定簇。利用表达hMPV F蛋白的重组杆状病毒感染粉纹夜蛾（Trichoplusia ni，Tn 5）昆虫细胞，建立了一种新的免疫荧光检测方法（Bac-F IFA）。采用Bac-F IFA检测了200份1个月至41岁日本人血清样本中hMPV F蛋白的免疫球蛋白G抗体。将结果与基于hMPV感染的LLC-MK2细胞的常规IFA（hMPV IFA）的结果进行比较。两种间接免疫荧光法测定的抗体滴度相关性良好（相关系数为0.88），两种间接免疫荧光法测定的血清反应性一致性为91%（kappa = 0.76）。对于200份血清样本中的192份，Bac-F IFA获得的滴度等于或高于hMPV IFA获得的滴度。这些结果表明Bac-F IFA比hMPV IFA更敏感，并且hMPV IFA检测到的大多数抗体与hMPV F蛋白反应。Bac-F IFA是一种比hMPV IFA更可靠、灵敏和特异的hMPV抗体检测方法。
Human metapneumovirus (hMPV) has recently been identified as an etiological agent of acute respiratory infections. The hMPV fusion (F) protein has been indicated to be a major antigenic determinant that mediates effective neutralization and protection against hMPV infection. We developed a new immunofluorescence assay (IFA) using Trichoplusia ni (Tn5) insect cells infected with a recombinant baculovirus-expressing hMPV F protein (Bac-F IFA). A total of 200 serum samples from Japanese people I month to 41 years old were tested for immunoglobulin G antibodies to hMPV F protein by Bac-F IFA. The results were compared with those of the conventional IFA based on hMPV-infected LLC-MK2 cells (hMPV IFA). The titers obtained by the two IFAs correlated well (correlation coefficient of 0.88), and the concordance of seroreactivities between the two IFAs was 91% (kappa = 0.76). For 192 of the 200 serum samples, the titers obtained by the Bac-F IFA were equal to or higher than those obtained by the hMPV IFA. These results indicated that the Bac-F IFA was more sensitive than the hMPV IFA and that the majority of the antibodies detected by the hMPV IFA reacted with the hMPV F protein. The Bac-F IFA is a more reliable, sensitive, and specific method for the detection of hMPV antibodies than is the hMPV IFA.