Intact parathyroid hormone: performance and clinical utility of an automated assay based on high-performance immunoaffinity chromatography and chemiluminescence detection.
The performance and clinical utility of an automated assay of intact parathyroid hormone (parathyrin, PTH) are evaluated. The method is based on the extraction of PTH from plasma by an HPLC column containing immobilized anti-(44-68 PTH) antibodies. The PTH retained is detected with a postcolumn reactor and use of anti-(1-34 PTH) chemiluminescent-labeled antibodies. The total cycle time of the assay is 6.5 min per injection after a 1-h incubation. The lower limit of detection for PTH in a 66-microL plasma sample was 0.5 pmol/L based on peak heights and 0.2 pmol/L based on peak areas. Mean analytical recovery for PTH added to plasma was 97%. The within-day precisions (CVs) for 4.2 and 30 pmol/L PTH plasma samples were 9.2% and 5.6% and the day-to-day precisions were 10.3% and 5.7%, respectively. No significant interferences from 1-34, 44-68, or 53-84 PTH fragments were noted, even at highly increased concentrations of fragments. The correlation of results with those of a manual assay of intact PTH was 0.97, and the results showed good agreement with disease state for patients with hypo- or hyperparathyroidism. The specificity of the assay for primary hyperparathyroidism was greater than 95%. We discuss the advantages (speed and quality control) of this approach over current immunoassays and the potential use of this method for detecting other analytes.
Hage
D S
DS
Department of Chemistry, University of Nebraska, Lincoln 68588-0304.
Taylor
B
B
Kao
P C
PC
D001322:Autoanalysis / Q000379:methods* / Q000706:statistics & numerical data; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D006934:Hypercalcemia / Q000097:blood / Q000209:etiology; D006961:Hyperparathyroidism / Q000097:blood; D007676:Kidney Failure, Chronic / Q000097:blood; D008163:Luminescent Measurements; D009369:Neoplasms / Q000150:complications; D010281:Parathyroid Hormone / Q000097:blood*; D010446:Peptide Fragments / Q000097:blood; D011786:Quality Control; D012016:Reference Values
D010281:Parathyroid Hormone; D010446:Peptide Fragments