Indigenous anti-hepatitis A virus IgM capture ELISA for the diagnosis of hepatitis A.
Anti-hepatitis A virus IgM capture ELISA was developed by using the reagents produced in the NIV laboratory. The major reagents of the assay were anti-human IgM antibody, hepatitis A virus (HAV) and anti-HAV IgG-horse radish peroxidase (HRP) conjugate. Of these, anti-human IgM antibodies were generated in rabbit against IgM secreted by human hybridoma clone(G3). HAV was derived from buffalo green money kidney cell line infected with HM-175 strain. Virus purified from the cell lysates was used for immunization of rabbits and guinea-pigs. There was very low anti-HAV response. A seropositive rhesus monkey was inoculated with monkey adapted strain of HAV to boost the anti-HAV antibody titre. Anti-HAV IgGs derived from hyperimmune sera of monkey and hepatitis A patient were conjugated with HRP. The preparations of conjugate--particularly human antibody--HRP conjugate yielded highly satisfactory results in anti-HAV capture ELISA. The assay appears to be specific, sensitive and quick and is useful in differentiating acute HAV infection from other acute infections caused by B, E and non-A non-B hepatitis viruses.
Chitambar
S D
SD
National Institute of Virology, Pune.
Murthy-Grewal
S
S
Bokil
M
M
Arankalle
V A
VA
Gore
M M
MM
Banerjee
K
K
D000818:Animals; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D006506:Hepatitis A / Q000175:diagnosis*; D035922:Hepatitis A Antibodies; D006508:Hepatitis Antibodies / Q000097:blood*; D006507:Hepatovirus / Q000276:immunology*; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood*; D011817:Rabbits
D035922:Hepatitis A Antibodies; D006508:Hepatitis Antibodies; D007075:Immunoglobulin M