Abstract:
:There is a lack of consensus among investigators who use a variety of immunoassay techniques (e.g., enzyme-linked immunosorbent assay [ELISA] and radioimmunoassay) regarding the protocols for describing and forming standard reference or calibration curves and interpolating serum antibody concentrations. This confounds the issue of detecting the presence or absence of parallelism between standard reference serum and serially diluted serum sample curves. These curves must be parallel to support the assumption that the antibody-binding characteristics are similar enough to allow the determination of antibody levels in the diluted serum sample. There is no universal and widely adopted strategy for assessing parallelism in bioassays, and without an assurance of parallelism, investigators are not able to calculate reliable estimates for antibody concentrations in serum samples. Furthermore, single-point (dilution) serum assays do not provide information related to parallelism and nonparallelism, and this, too, may lead to considerable error when calculating antibody concentrations. When assay methodology, technique, and precision improve to the extent that standard reference serum and serially diluted serum sample curves are fit with little error, standard analysis of variance techniques are overly sensitive to negligible departures from parallelism. We present a series of guidelines that compose a protocol for assessing parallelism between bioassay dilution curves that are applicable to data derived from ELISAs. These criteria should be applicable, with minor modifications, to most immunoassay experimental situations and, most importantly, are not dependent on the mathematical model used to form the standard reference curve. These guidelines have evolved in our laboratories over the past 4 years during the performance of thousands of ELISAs for antibodies to the capsular polysaccharides of Neisseria meningitidis groups A and C and Haemophilus influenzae type b.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Plikaytis BD,Holder PF,Pais LB,Maslanka SE,Gheesling LL,Carlone GMdoi
10.1128/JCM.32.10.2441-2447.1994subject
Has Abstractpub_date
1994-10-01 00:00:00pages
2441-7issue
10eissn
0095-1137issn
1098-660Xjournal_volume
32pub_type
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.25.5.840-844.1987
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doi:10.1128/JCM.32.3.828-831.1994
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