Type- and subtype-specific detection of influenza viruses in clinical specimens by rapid culture assay.
A rapid culture assay which allows for the simultaneous typing and subtyping of currently circulating influenza A(H1N1), A(H3N2), and B viruses in clinical specimens was developed. Pools of monoclonal antibodies (MAbs) against influenza A and B viruses and MAbs HA1-71 and HA2-76, obtained by immunizing mice with the denatured hemagglutinin subfragments HA1 and HA2 of influenza virus A/Victoria/3/75, were used for immunoperoxidase staining of antigens in infected MDCK cells. MAb HA1-71 reacted exclusively with influenza A viruses of the H3 subtype, while MAb HA2-76 reacted with subtypes H1, H3, H4, H6, H8, H9, H10, H11, and H12, as determined with 78 human, 4 swine, and 10 avian influenza virus reference strains subtyped by the hemagglutination inhibition test. To determine if the technique can be used as a rapid diagnostic test, 263 known influenza virus-positive frozen nasal or throat swabs were inoculated into MDCK cells. After an overnight incubation, the cells were fixed and viral antigens were detected by immunoperoxidase staining. Influenza A viruses of the H1 and H3 subtypes were detected in 31 and 113 specimens, respectively. The subtypes of 10 influenza A virus-positive specimens could not be determined because they contained too little virus. Influenza B viruses were detected in 84 specimens, and 25 specimens were negative. We conclude that this assay is a rapid, convenient, non-labor-intensive, and relatively inexpensive test for detecting, typing, and subtyping influenza viruses in clinical specimens.
Ziegler
T
T
Influenza Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333.
Hall
H
H
Sánchez-Fauquier
A
A
Gamble
W C
WC
Cox
N J
NJ
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D002460:Cell Line; D004196:Disease Outbreaks; D004285:Dogs; D005069:Evaluation Studies as Topic; D005188:False Negative Reactions; D006801:Humans; D007124:Immunoenzyme Techniques; D009980:Influenza A virus / Q000145:classification / Q000276:immunology / Q000302:isolation & purification; D009981:Influenza B virus / Q000145:classification / Q000276:immunology / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology*; D051379:Mice; D009975:Orthomyxoviridae / Q000145:classification* / Q000276:immunology / Q000302:isolation & purification*; D012703:Serotyping; D013997:Time Factors; D014776:Virus Cultivation / Q000379:methods*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral