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Impact of old and current immunoassays on the 1 mg overnight dexamethasone suppression test: comparison with LC-MS/MS
Laura Rotolo1,2, Greta Galante2,3, Kimberly Coscia1,2
1Division of Endocrinology and Diabetes Prevention and Care, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna 40138, Italy.
Insights
Two common immunoassays underdetect hypercortisolism compared to LC-MS/MS, necessitating method-specific cutoffs for accurate diagnosis. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is preferred for hypercortisolism testing.
Area of Science:
- Clinical Chemistry
- Endocrinology
- Diagnostic Assays
Background:
- Hypercortisolism diagnosis relies on accurate cortisol measurements.
- The performance of immunoassays in dexamethasone suppression test (DST) settings is not well-established.
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the recommended reference method.
Purpose of the Study:
- To evaluate the effectiveness of two immunoassays (Elecsys gen I and Access) in detecting hypercortisolism.
- To compare immunoassay performance against LC-MS/MS.
- To assess immunoassay analytical performance in basal and post-DST conditions.
Main Methods:
- Cortisol measurement in post-DST sera from 260 (Elecsys gen I) and 217 (Access) patients using immunoassays and LC-MS/MS.
- Estimation of hypercortisolism rates using a 50 nmol/L cutoff.
- Generation of method-specific cutoffs for >95% sensitivity and >80% specificity.
- Comparison of cortisol measurements in basal and post-DST samples.
Main Results:
- Both immunoassays showed lower hypercortisolism detection rates than LC-MS/MS, especially in adrenal adenoma patients.
- Immunoassays yielded potential false negatives (6.4-6.9%) and false positives (3.8% for Elecsys gen I).
- Optimal cutoffs were 41 nmol/L for Elecsys gen I and 33 nmol/L for Access.
- Immunoassays showed significant deviations from LC-MS/MS in basal and post-DST samples, with noted sex differences.
Conclusions:
- Immunoassays significantly underdetect hypercortisolism, supporting the need for method-specific cutoffs.
- Immunoassay performance varies between basal and post-DST conditions.
- LC-MS/MS methods are recommended for accurate hypercortisolism assessment.
Objective:
The influence of immunoassay performance in hypercortisolism and dexamethasone suppression test (DST) settings was scarcely investigated. We evaluated the effectiveness of 2 immunoassays in detecting hypercortisolism compared to recommended liquid chromatography-tandem mass spectrometry (LC-MS/MS), and compared immunoassay analytical performance in basal and post-DST conditions.
Methods:
We measured cortisol in post-DST sera of patients with suspected hypercortisolism or adrenal incidentalomas by Elecsys gen I (n = 260), and by Access (n = 217). All samples were also measured by a validated LC-MS/MS method. We estimated hypercortisolism rate according to the established 50 nmol/L cutoff, and generated immunoassay-specific cutoffs providing >95% sensitivity and >80% specificity. Finally, we compared cortisol measurements in basal and post-DST samples.
Results:
Using the 50 nmol/L cutoff, both immunoassays detected lower rates of hypercortisolism compared with LC-MS/MS, particularly in patients with adrenal adenomas (P < .050). Elecsys gen I and Access determined 6.9% and 6.4% possible false negatives, respectively. Elecsys gen I also caused 3.8% possible false positives. Optimal cutoff was 41 nmol/L for Elecsys gen I (sensitivity: 97.7%; specificity: 80.8%), and 33 nmol/L for Access (sensitivity: 97.5%; specificity: 78.3%). In basal and post-DST samples, Elecsys gen I overestimated by 32.5% and 6.1%, whereas Access underestimated by -4.7% and -5.9% compared to LC-MS/MS cortisol measurements, respectively. Sex differences in method deviations were noted.
Conclusions:
Both immunoassays demonstrated remarkable underdetection of hypercortisolism, suggesting the application of a method-specific cutoff. Immunoassay performance may not be uniform in basal and post-DST conditions and should be purposely examined. Accurate LC-MS/MS methods should be preferred in hypercortisolism settings.
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