Enzyme-linked immunosorbent assay for measurement of antibody against cytomegalovirus and rubella virus in a single serum dilution.

Abstract

SUMMARY Enzyme-linked immunosorbent assays (ELISA) were developed for quantifying cytomegalovirus (CMV) and rubella antibodies using a single serum dilution (1/800) in conjunction with a standard curve. A near linear relation was found between the logarithms of absorbance values of sera at a dilution of 1/800 and the titres as determined by an end point dilution ELISA. The reproducibility of the single dilution ELISA was good; the within-test coefficients of variation averaged 7.5 % for CMV antibody and 12-4 % for rubella antibody. A close correlation was found between ELISA and complement-fixing (CF) antibody titres to CMV and between ELISA and haemagglutination-inhibition (HI) antibody titres to rubella virus. The titres in ELISA were 200 to 1000 times higher than in CF for CMV and 50 to 100 times higher than in HI for rubella virus.Many reports have described the application of the enzyme-linked immunosorbent assay (ELISA) for laboratory diagnosis of infectious diseases.' 2 With respect to sensitivity, specificity, reproducibility, and ease of performance, ELISA has distinct advantages over existing techniques for determining antibody titres to different aetiological agents including cytomegalovirus (CMV)3-6 and rubella virus.3 7-10 CMV and rubella antibody levels in ELISA have been determined by examining serial serum dilutions4 6 7 10 or a single serum dilution.3 5 8 It is apparent from these studies that there is a need to standardise methods to distinguiish between specific and nonspecific reactivity.We report an ELISA procedure which permits a precise and reproducible quantification of CMV and rubella antibodies by examination of a single serum dilution in conjunction with a standard curve. The sensitivity of the test is compared with that of complement fixation (CF) for CMV and haemagglutination inhibition (HI) for rubella virus. Material and methods ANTIGENS

Elisa test for the serodiagnosis of Akabane virus infection in cattle

“…This makes it possible to determine antibody titre by assay of a single serum dilution in this range when carried out in conjunction with a relevant standard curve (Figs 2 and 3). However when sera are examined at lower dilutions endpoint titration curves for antibodies of either class should be performed for each serum sample (Van Loon et al, 1981). ELISA was found to be more sensitive than SN (Table II).…”

Frequency distributions in rainbow trout populations of absorbance values from an ELISA for Vibrio anguillarum antibodies

“…A ?g critical inltial step in this process is to investigate the variation of OD in negative populations and in diseased a n d immunized populations. Workers in human and veterinary medicine have found the distribution of ELISA values in non-diseased populations to follow a normal pattern (van Loon et al 1981) or to have a positive skew (de Savigny et al 1979, Gillis et al 1984. Quantitative information on the distribution in diseased populations is less available.…”

“…Some workers have used arbitrarily determined fixed levels (Bortz et al 1984, CossariniDunier 1985. It is preferable to base cut-off levels on the frequency distribution of the OD of negative sera (van Loon et al 1981). Kodoma et al (1985) and Thorburn (1986) set the minimum positive OD at the mean OD of samples collected from non-exposed populations O Inter-Research/Printed in F. R. Germany plus 2 or 3 standard deviations of the mean.…”

Quantitative and qualitative detection of cytomegalovirus‐specific antibodies using two types of enzyme‐linked immunosorbent assay

“…These techniques are being replaced by the more sensitive techniques of radioimmunoassay [Knez et al, 1976; Janowski et al, 198QI-and enzyme-linked immunosorbent assay (ELISA) [Booth et al, 1979;Sarov et al, 1980;Van Loon et al, 1981; Kiefer et al, 19831. Both these techniques have comparable sensitivity and specificity in the detection of CMV-specific antibody [Booth et al, 19821.…”