Detection of Hendra viral IgG antibody in animal sera is useful for surveillance following a virus outbreak. The commonly used enzyme-linked immunosorbent assay and fluorescence-based Luminex assay typically consist of three steps and take at least several hours to complete. We have simplified the procedure to two steps in an effort to develop a rapid procedure for IgG antibody, but not IgM antibody, detection. This is achieved by conjugating the fluorescence label R-phycoerythrin directly onto the IgG binding protein Protein G. The use of magnetic nanoparticles, due to their large specific surface area, has helped reduce each of the binding steps to 20 min. As a result, the whole assay can be completed in 60 min. We also demonstrate a method to quickly estimate IgG antibody titres by assaying the sera at only two dilutions (i.e. 1:20 and 1:1000) and using the fluorescence ratio at these dilutions as an indicator of antibody titre. The results of this approach correlated well with the well-regarded serum neutralization test in virus antibody assays. This protocol reported here can be adopted in Luminex assays, fluorescence-linked immunosorbent assays and assays on microfluidics platforms for rapid antibody surveillance of Hendra and other viruses. (C) 2015 Elsevier B.V. All rights reserved.
机构:
KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, SwedenKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Ayoglu, Burcu
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Szarka, Eszter
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Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Szarka, Eszter
;
Huber, Krisztina
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Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Huber, Krisztina
;
Orosz, Anita
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Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Orosz, Anita
;
Babos, Fruzsina
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MTA ELTE Res Grp Peptide Chem, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Babos, Fruzsina
;
Magyar, Anna
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h-index: 0
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MTA ELTE Res Grp Peptide Chem, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Magyar, Anna
;
Hudecz, Ferenc
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机构:
MTA ELTE Res Grp Peptide Chem, Budapest, Hungary
Eotvos Lorand Univ, Dept Organ Chem, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Hudecz, Ferenc
;
Rojkovich, Bernadette
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Polyclin Hospitaller Bros St John God, Dept Rheumatol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Rojkovich, Bernadette
;
Gati, Tamas
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Polyclin Hospitaller Bros St John God, Dept Rheumatol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Gati, Tamas
;
Nagy, Gyoergy
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Polyclin Hospitaller Bros St John God, Dept Rheumatol, Budapest, Hungary
Semmelweis Univ, Sch Med, Dept Genet Cell & Immunobiol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Nagy, Gyoergy
;
Schwenk, Jochen M.
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KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, SwedenKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Schwenk, Jochen M.
;
Sarmay, Gabriella
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Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Sarmay, Gabriella
;
Prechl, Jozsef
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h-index: 0
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MTA ELTE Immunol Res Grp, Budapest, Hungary
Diagnosticum Ltd, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Prechl, Jozsef
;
Nilsson, Peter
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KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, SwedenKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Nilsson, Peter
;
Papp, Krisztian
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KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
MTA ELTE Immunol Res Grp, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
机构:
KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, SwedenKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Ayoglu, Burcu
;
Szarka, Eszter
论文数: 0引用数: 0
h-index: 0
机构:
Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Szarka, Eszter
;
Huber, Krisztina
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h-index: 0
机构:
Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Huber, Krisztina
;
Orosz, Anita
论文数: 0引用数: 0
h-index: 0
机构:
Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Orosz, Anita
;
Babos, Fruzsina
论文数: 0引用数: 0
h-index: 0
机构:
MTA ELTE Res Grp Peptide Chem, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Babos, Fruzsina
;
Magyar, Anna
论文数: 0引用数: 0
h-index: 0
机构:
MTA ELTE Res Grp Peptide Chem, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Magyar, Anna
;
Hudecz, Ferenc
论文数: 0引用数: 0
h-index: 0
机构:
MTA ELTE Res Grp Peptide Chem, Budapest, Hungary
Eotvos Lorand Univ, Dept Organ Chem, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Hudecz, Ferenc
;
Rojkovich, Bernadette
论文数: 0引用数: 0
h-index: 0
机构:
Polyclin Hospitaller Bros St John God, Dept Rheumatol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Rojkovich, Bernadette
;
Gati, Tamas
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h-index: 0
机构:
Polyclin Hospitaller Bros St John God, Dept Rheumatol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Gati, Tamas
;
Nagy, Gyoergy
论文数: 0引用数: 0
h-index: 0
机构:
Polyclin Hospitaller Bros St John God, Dept Rheumatol, Budapest, Hungary
Semmelweis Univ, Sch Med, Dept Genet Cell & Immunobiol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Nagy, Gyoergy
;
Schwenk, Jochen M.
论文数: 0引用数: 0
h-index: 0
机构:
KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, SwedenKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Schwenk, Jochen M.
;
Sarmay, Gabriella
论文数: 0引用数: 0
h-index: 0
机构:
Eotvos Lorand Univ, Dept Immunol, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Sarmay, Gabriella
;
Prechl, Jozsef
论文数: 0引用数: 0
h-index: 0
机构:
MTA ELTE Immunol Res Grp, Budapest, Hungary
Diagnosticum Ltd, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Prechl, Jozsef
;
Nilsson, Peter
论文数: 0引用数: 0
h-index: 0
机构:
KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, SwedenKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
Nilsson, Peter
;
Papp, Krisztian
论文数: 0引用数: 0
h-index: 0
机构:
KTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden
MTA ELTE Immunol Res Grp, Budapest, HungaryKTH Royal Inst Technol, Sch Biotechnol, SciLifeLab, Stockholm, Sweden