title: Detection of methicillin-resistant Staphylococcus aureus using a specific anti-PBP2a chicken IgY antibody.
abstract: Immunodetection of methicillin-resistant Staphylococcus aureus (MRSA) by conventional methods employing mammalian immunoglobulins has unknown detection limits, and often yields false-positive results because of the presence of S. aureus protein A, which binds the Fc region of mammalian IgG. In this study, a new PBP2a-specific chicken IgY antibody was developed in inbred and conventional chickens, and used for the detection of MRSA using whole cell lysate samples. Our results showed that this chicken IgY antibody minimized the side effects of protein A. Moreover, enzyme-linked immunosorbent assay and immunochromatography systems were used with a monoclonal and polyclonal anti-PBP2a IgY antibody, clearly differentiating MRSA from methicillin-sensitive S. aureus and other methicillin-sensitive Staphylococcus spp. The detection limit of the immunochromatography was 10(8) colony-forming units; therefore, 1 colony on an agar plate was adequate to distinguish MRSA from non-MRSA. The specificity and sensitivity of this assay were almost similar to that of a commercially available latex agglutination test; however, the procedure used in this study was less complicated. The entire detection procedure, including sample preparation, takes only 20 min and does not require special equipment. Therefore, the use of this IgY antibody as a new tool for the detection of MRSA is highly recommended.
authors.affiliation: Department of Bacteriology, Graduate School of Medicine, Nagoya University, Nagoya 466-8550, Japan. yakeiko＠med.nagoya-u.ac.jp
authors.forename: Keiko
authors.initials: K
authors.lastname: Yamada
authors.forename: Jin
authors.initials: J
authors.lastname: Wanchun
authors.forename: Teruko
authors.initials: T
authors.lastname: Ohkura
authors.forename: Atsushi
authors.initials: A
authors.lastname: Murai
authors.forename: Reina
authors.initials: R
authors.lastname: Hayakawa
authors.forename: Keiji
authors.initials: K
authors.lastname: Kinoshita
authors.forename: Makoto
authors.initials: M
authors.lastname: Mizutani
authors.forename: Akira
authors.initials: A
authors.lastname: Okamoto
authors.forename: Takao
authors.initials: T
authors.lastname: Namikawa
authors.forename: Michio
authors.initials: M
authors.lastname: Ohta
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial; D001431:Bacteriological Techniques / Q000379:methods*; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007136:Immunoglobulins; D055624:Methicillin-Resistant Staphylococcus aureus / Q000737:chemistry / Q000302:isolation & purification*; D046915:Penicillin-Binding Proteins / Q000032:analysis*; D010453:Peptide Synthases / Q000032:analysis*; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000382:microbiology*; D013997:Time Factors
chemical_list: D000907:Antibodies, Bacterial; C016512:IgY; D007136:Immunoglobulins; D046915:Penicillin-Binding Proteins; C478699:penicillin-binding protein 2a, Streptococcus; D010453:Peptide Synthases