title: Adaptation of the Diazyme Direct Enzymatic HbA1c Assay for a microplate reader at room temperature.
abstract: BACKGROUND
  Measurement of hemoglobin A(1c) (HbA(1c)) is a key diagnostic criterion and a key parameter for the follow-up of the treatment of diabetes mellitus. Typically, immunochemical assays of HbA(1c) are performed in clinical chemistry analyzers. In this study, we applied the HbA(1c) assay on a microplate reader at room temperature.
  
  
  METHODS
  HbA(1c) samples were measured using the Direct Enzymatic HbA(1c) Assay from Diazyme Laboratories (Poway, CA, USA) using a Plate Chameleon Microplate Reader (Hidex Co., Turku, Finland) according to the manufacturer's protocol and a modification of the method to room temperature. The Tosoh G7 HPLC method for HbA(1c) (Tosoh Co., Tokyo, Japan) was used as a comparative method.
  
  
  RESULTS
  There was good correlation of HbA(1c) results when the assay was performed at room temperature (+22°C) compared with that at +37°C (r=0.987). The modified method was linear over the HbA(1c) range 4%-14%. Analysis of HbA(1c) results from 50 blood samples by the modified method showed good agreement with the HPLC method (r=0.990).
  
  
  CONCLUSIONS
  The modified Diazyme Direct Enzymatic HbA(1c) Assay™ appears to work as good at +22°C as that performed according to manufacturer's protocol at +37°C.
authors.affiliation: Kuopio Research Institute of Exercise Medicine, Kuopio, Finland.
authors.forename: Ilkka
authors.initials: I
authors.lastname: Penttilä
authors.forename: Karri
authors.initials: K
authors.lastname: Penttilä
authors.forename: Toivo
authors.initials: T
authors.lastname: Halonen
authors.forename: Kari
authors.initials: K
authors.lastname: Pulkki
authors.forename: Jukka
authors.initials: J
authors.lastname: Törrönen
authors.forename: Rainer
authors.initials: R
authors.lastname: Rauramaa
mesh_terms: D001774:Blood Chemical Analysis / Q000379:methods*; D003920:Diabetes Mellitus / Q000097:blood; D057075:Enzyme Assays; D006442:Glycated Hemoglobin / Q000032:analysis*; D006801:Humans; D007118:Immunoassay; D009195:Peroxidase / Q000378:metabolism; D013696:Temperature
chemical_list: D006442:Glycated Hemoglobin; D009195:Peroxidase; C517652:hemoglobin A1c protein, human