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.
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.
Uchida
Eriko
E
Division of Cellular and Gene Therapy Products, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya, Tokyo 158-8501, Japan.
Kogi
Mieko
M
Oshizawa
Tadashi
T
Furuta
Birei
B
Satoh
Koei
K
Iwata
Akiko
A
Murata
Mitsuhiro
M
Hikata
Mikio
M
Yamaguchi
Teruhide
T
D000936:Antigen-Antibody Complex; D016174:Hepacivirus / Q000276:immunology / Q000302:isolation & purification*; D017957:Hepatitis A Virus, Human / Q000276:immunology / Q000302:isolation & purification*; D006515:Hepatitis B virus / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008863:Microspheres; D011094:Polyethyleneimine; D016133:Polymerase Chain Reaction
D000936:Antigen-Antibody Complex; D011094:Polyethyleneimine