Optimization of the virus concentration method using polyethyleneimine-conjugated magnetic beads and its application to the detection of human hepatitis A, B and C viruses
Optimization of the virus concentration method using polyethyleneimine-conjugated magnetic beads and its application to the detection of human hepatitis A, B and C viruses
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DOI：
10.1016/j.jviromet.2007.02.014
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发表时间：
2007-07-01
影响因子：
3.1
通讯作者：
Yamaguchi, Teruhide
中科院分区：
文献类型：
--
作者：
Uchida, Eriko;Kogi, Mieko;Yamaguchi, Teruhide
为了提高聚合酶链反应（PCR）和逆转录PCR（RT-PCR）检测病毒的灵敏度，我们在前期研究中建立了一种利用聚乙烯亚胺（PEI）标记的磁珠富集病毒的新方法。然而，有几种病毒不能用这种方法浓缩。本文对病毒浓缩条件进行了优化，以更有效地浓缩多种病毒。PEI珠吸附病毒的效率比其他阳离子聚合物，并在弱酸性条件下获得最佳的病毒浓度。质谱分析表明，几种血清蛋白，如补体3型，补体4型和免疫球蛋白M（IgM），共吸附的PEI珠，这表明珠吸附病毒不仅通过直接吸附，而且通过免疫复合物的形成。PEI珠不能直接吸附的脊髓灰质炎病毒可以通过免疫复合物的形成被珠浓缩，这一结果证实了这一假设。另一方面，甲肝（HAV）和丙肝（HCV）病毒几乎完全被PEI珠直接吸附。与脊髓灰质炎病毒一样，加入抗HBV IgM可有效浓缩B型肝炎病毒（HBV）。总之，使用PEI珠的病毒浓缩是一种有用的方法，可用于浓缩各种病毒，并可用于提高HAV、HBV和HCV检测的灵敏度。(C)2007 Elsevier B. V.保留所有权利。
To enhance the sensitivity of virus detection by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR), a novel virus concentration method using polyethyleneimine (PEI)-conjugated magnetic beads was developed in our previous study. However, several viruses could not be concentrated by this method. In this paper, the conditions of virus concentration were optimized to concentrate a wide range of viruses more efficiently. The PEI beads adsorbed viruses more efficiently than other cationic polymers, and the optimum virus concentration was obtained under weak acidic conditions. Mass spectrometric analysis revealed that several serum proteins, such as complement type 3, complement type 4 and immunoglobulin M (IgM), were co-adsorbed by the PEI beads, suggesting that the beads may adsorb viruses not only by direct adsorption, but also via immune complex formation. This hypothesis was confirmed by the result that poliovirus, which PEI beads could not adsorb directly, could be concentrated by the beads via immune complex formation. On the other hand, hepatitis A (HAV) and hepatitis C (HCV) viruses were adsorbed directly by PEI beads almost completely. Like poliovirus, hepatitis B virus (HBV) was concentrated efficiently by the addition of anti-HBV IgM. In conclusion, virus concentration using PEI beads is a useful method to concentrate a wide range of viruses and can be used to enhance the sensitivity of detection of HAV, HBV and HCV. (C) 2007 Elsevier B.V. All rights reserved.