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Clinical Chemistry 48: 613-621, 2002;
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(Clinical Chemistry. 2002;48:613-621.)
© 2002 American Association for Clinical Chemistry, Inc.

Immunometric Assay Interference

Incidence and Prevention

Johan Bjerner1a, Kjell Nustad1, Lars F. Norum1, Kari Hauge Olsen1 and Ole P. Børmer1

1 Central Laboratory, Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway.

aAuthor for correspondence. Fax 47-22-730725; e-mail johan.bjerner{at}klinmed.uio.no.

Background: The primary aim of the study was to reduce interference in an in-house two-site, two-step immunometric assay.

Methods: In the running laboratory routine, 11 261 samples were tested with a carcinoembryonic antigen (CEA) assay with bovine immunoglobulin but no murine immunoglobulins in the buffer, in parallel to our routine CEA assay, using 15 mg/L heat-treated nonspecific murine immunoglobulin (MAK33) in the buffer and with the Fc fragments removed from the capture antibody.

Results: The frequency of interference was estimated to be 4.0% (95% confidence interval, 3.3–4.7%). The addition of 15 mg/L native MAK33 had little effect (frequency, 3.9%; 95% confidence interval, 3.2–4.6%), whereas adding 15 mg/L heat-treated MAK33 reduced interference to 0.86% (0.61–1.12%), and adding 50 mg/L reduced it further to 0.06% (0–0.13%). Removing the Fc fragments by itself reduced interference to 0.10% (0.02–0.19%). There were no statistically significant differences for age (P <0.23) or gender (P <0.40) between patients with interference (n = 210) and a randomly selected interference-negative control group (n = 186). Interference was not constant in patients: 15 of 25 individuals positive for interference and with four or more samples screened for interference had an interference-negative sample either before or after the peak of interference.

Conclusions: In a two-site, two-step immunometric assay using mouse monoclonal antibodies, use of heat-treated nonspecific murine immunoglobulin in the buffer or removal of the Fc fragment from the capture antibody could improve performance.




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