Thrombin-mediated degradation of parathyroid hormone in serum tubes

Intact parathyroid hormone (PTH) tests are frequently sandwich immunoassays. Enzymes that cleave PTH may cause falsely lower PTH results. The objective of this study was to determine whether bovine thrombin in Becton Dickinson (BD) Vacutainer rapid serum tubes™ (RSTs) may lead to PTH results that are lower than in plasma separator tube™ (PST) or serum separator tube™ (SST) collections.

Tubes of blood (PST, SST, and RST) were collected from donors. PTH concentrations were measured on a Roche Cobas e602 analyzer in aliquots held at room temperature or 4°C across time. Instrument comparison studies were also conducted on an Abbott Architect i1000SR and a Siemens Immulite 2000 XPi. Previously collected serum specimens were also incubated in exogenous bovine thrombin, the direct thrombin inhibitor hirudin, or both. Freshly collected RST specimens were also spiked with hirudin after clotting and centrifugation.

Significant decreases in PTH degradation rate constants were observed according to tube type, with degradation rates faster in RSTs than SSTs, and SSTs faster than PSTs. PTH degradation rate was temperature dependent. PTH decreases induced by exogenous bovine thrombin, as well as endogenous human thrombin, were reduced by hirudin.

Bovine thrombin is responsible for the decrease in PTH results observed in RSTs. Endogenous human thrombin, activated during clot formation, is likely responsible for the smaller decreases observed in non-RST sera versus plasma.