Premium
Detection of antibodies against human metapneumovirus by western blot using recombinant nucleocapsid and matrix proteins
Author(s) -
Ishiguro Nobuhisa,
Ebihara Takashi,
Endo Rika,
Ma Xiaoming,
Kawai Eri,
Ishiko Hiroaki,
Kikuta Hideaki
Publication year - 2006
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.20667
Subject(s) - human metapneumovirus , recombinant dna , western blot , virology , antibody , titer , viral matrix protein , biology , microbiology and biotechnology , immunofluorescence , immunogenicity , antigen , antibody titer , virus , immunology , respiratory system , respiratory tract infections , biochemistry , gene , anatomy
Abstract Detection of antibodies against individual proteins of human metapneumovirus (hMPV) is important in the analysis of immune responses to hMPV. Specific antibodies against nucleocapsid (N) and matrix (M) proteins in 97 serum samples were tested by Western blot using recombinant N and M proteins of hMPV expressed in Escherichia coli . The results were compared with those of immunofluorescence assays (IFAs) based on hMPV‐infected LLC‐MK2 cells, which expressed the whole hMPV proteins. Thirty (61.2%) and 31 (63.3%) of 49 serum samples with titers of >/=1:160 by IFA reacted with N and M proteins, respectively. Only 2 (4.2%) of 11 serum samples with titers of 1:80 by IFA reacted with N and M proteins. Antibodies against N and M proteins were not detected in 37 serum samples with titers of <1:40 by IFA. These results indicate that the antibodies against N and M proteins are highly specific (100%) but less sensitive (42.1%, N protein; 40.8%, M protein) than those against whole proteins of hMPV detected by IFA. The reactivity of sera with the recombinant N protein and that with the recombinant M protein correlated well (correlation coefficient of 0.79), and the concordance of reactivities was 91% (κ = 0.79). In summary, both recombinant N and M proteins of hMPV were antigenic, and the responses to N and M protein varied among patients. Therefore, Western blot using N and M proteins provide a useful tool for analysis of immune responses to hMPV. J. Med. Virol. 78:1091–1095, 2006. © 2006 Wiley‐Liss, Inc.