Comparison of two chromogranin A assays and investigation of nonlinear specimens.

Background: As a marker for functional and non-functional neuroendocrine tumors, serum chromogranin A (CgA) concentrations have shown value for detecting and monitoring disease. Here we describe a comparison between an established micro-titer plate assay (Cisbio CgA ELISA) and an analyzer-based assay (B·R·A·H·M·S CgA II KRYPTOR). Reference limits were established along with a performance evaluation of the KRYPTOR assay. Nonlinearity observed in approximately 0.03% of patients was also investigated.

Methods: Samples were tested according to kit manufacturer's protocols. Reference limits were established for both assays testing the same cohort of healthy volunteers. Potential causes of nonlinearity investigated were HAMA, macromolecule effects and elevated serum creatinine.

Results: KRYPTOR vs. Cisbio: slope=0.692, y-intercept=-40.0 (r=0.967, n=186). Upper reference limits were 160 and 103 ng/mL for the Cisbio and KRYPTOR assays, respectively. Linearity: slope=1.012 (r=0.998) with 95.0-105.5% recoveries. Precision: repeatability ≤2.4%, within-laboratory ≤3.1% (79 and 738 ng/mL). Limit of detection: 8 ng/mL. Strong nonlinear specimens (n=6) retested for HAMA interference generated differences (block-no block) ranging -3.2-4.2%. Polyethylene glycol precipitation recoveries ranged from 157 to >5714% for affected specimens versus 71-79% for normal specimens. Eight of 14 nonlinear specimens (57%) had elevated serum creatinine results (>1.20 mg/dL).

Conclusions: The CgA II KRYPTOR assay performs acceptably for quantifying CgA in human serum. While adequate correlation is observed against the Cisbio ELISA, there is significant disagreement overall. Efforts to identify a cause of the nonlinearity observed in a small percentage of patients were inconclusive, but neither HAMA interference, macromolecule effects nor renal failure appear as major factors.