Determination of immunoglobulin M antibodies against hepatitis B core antigen and hepatitis A virus by reorienting sucrose gradient high-speed centrifugation for diagnosis of acute viral-hepatitis

Abstract

Immunoglobulin M (IgM) antibodies against hepatitis B core antigen (anti-HBc) and hepatitis A virus (anti-HAV) were determined in 41 cases of acute viral hepatitis. In sera positive for anti-HBc or anti-HAV, IgM was separated from IgG by reorienting sucrose gradient high-speed centrifugation, and the IgG- and IgM-containing serum fractions were tested for the presence of specific antibody by radioimmunoassay. At the onset of illness, 4 of the 41 cases were classified as hepatitis A, 31 were hepatitis B, and 6 were non-A, non-B hepatitis, based on the results of these tests and of assays for hepatitis B surface antigen and antibody and hepatitis B e antigen and antibody. Fourteen of these 41 patients (34%) required IgM anti-HBc or IgM anti-HAV testing or both for appropriate classification. IgM anti-HBc persisted for at least 7 weeks after onset but no longer than 17 weeks in all patients tested with transient hepatitis B surface antigen-positive acute hepatitis. IgM anti-HAV persisted up to but not longer than 62 days in the patients with hepatitis A. Therefore, IgM anti-HBc and IgM anti-HAV determinants are valuable tools for the differential diagnosis of acute A, B, and non-A, non-B hepatitis.

Diagnostic Significance of Anti‐HBcIgM Prevalence Related to Symptoms in Canadian Patients Acutely or Chronically Infected With Hepatitis B Virus

“…The. use of staphylococcal protein A or SDG centrifugation combined with RIA for total anti-HBc or electrophoresis showed anti-HBcIgM disappearing from sera of acutely infected patients between 2 and 17 months after onset of symptoms [Cohen, 1978;Hawkes et al, 1980;Cappel et al, 1981;Swenson et al, 1981;Wang et al, 19841. Most of the solid phase enzyme-or radioimmunoassays have reported declines in the anti-HBcIgM titres in acutely infected patients by 6 months, with some persisting at low levels for years after symptoms had resolved [Aldershvile and Nielsen, 1980;Gerlich et al, 1980;Kryger et al, 1981a, b;Lemon et al, 1981;Roggerdorf et al, 1981;Chau et al, 19831.…”

“…Although the detection of serum hepatitis B surface antigen (HBsAg) has enabled the identification of acute and chronic infections, it was recognized in the late 1970s that IgM class antibodies to the hepatitis B core (HBc) antigen might provide further understanding of these different forms of virus infection. Some of these studies separated IgM from the rest of the immunoglobulins by treating serum with staphylococcal Cowan strain protein A or by employing sucrose density gradient (SDG) centrifugation with subsequent measurement of the fractions in electrophoresis or agglutination assays [Cohen, 1978;Supran et al, 19801 or solid phase radioimmunoassay (RIA) for the measurement of total antibody to HBc [Hawkes et al, 1980;Cappel et al, 1981;Swenson et al, 1981;Wang et al, 1984;Sampliner and Schable, 19841. There are several reports describing and evaluating solid phase capture antibody assays for measuring anti-HBcIgM by RIA [Tedder and Wilson-Croom, 1980;Lemon et al, 1981;Mortimer et al, 1981;Feinman et al, 1982;Banninger et al, 1983;Chau et al, 1983;Shimizu et al, 1983;Sjogren and Lemon, 1983;Sjogren and Hoofnagle, 19851 or by enzyme immunoassay (EIA) [Gerlich and Luer, 1979;Aldershvile and Nielsen, 1980;Gerlich et al, 1980Kryger et al, 1981a, b, 1982Roggerdorf et al, 1981;Goudeau et al, 1983;Perrillo et al, 1983;Coltorti et al, 1984;Nowicki et al, 1984;Papaevangelou et al, 1984, Gordon et al, 1985Hwang et al, 1985;van Diutzhuijsen et al, 19851, and novel tests combining RIA and EIA (Angarano et al, 1984) or enzyme-labeled antigen [Vilja et al, 19851 have appeared.…”