本试剂盒应用双抗体夹心酶标免疫分析法测定标本中HbA1c水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入HbA1c、生物素化的抗人HbA1c抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的HbA1c呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
This immunoassay kit allows for the specific measurement of human HbA1c concentrations in serum and plasma.
Introduction
hemoglobin, the compound in the red blood cells that transports oxygen. Hemoglobin occurs in several variants; the one which composes about 90% of the total is known as hemoglobin A. A1c is a specific subtype of hemoglobin A. The 1 is actually a subscript to the A, and the c is a subscript to the 1. "Hemoglobin" is also spelled "haemoglobin", depending on your geographic allegiance. Glucose binds slowly to hemoglobin A, forming the A1c subtype. The reverse reaction, or decomposition, proceeds relatively slowly, so any buildup persists for roughly 4 weeks. Because of the reverse reaction, the actual HbA1c level is strongly weighted toward the present. Some of the HbA1c is also removed when erythrocytes (red blood cells) are recycled after their normal lifetime of about 90-120 days. These factors combine so that the HbA1c level represents the average bG level of approximately the past 4 weeks, strongly weighted toward the most recent 2 weeks. It is almost entirely insensitive to bG levels more than 4 weeks previous. In non-diabetic persons, the formation, decomposition and destruction of HbA1c reach a steady state with about 3.0% to 6.5% of the hemoglobin being the A1c subtype. Most diabetic individuals have a higher average bG level than non-diabetics, resulting in a higher HbA1c level. The actual HbA1c level can be used as an indicator of the average recent bG level. This in turn indicates the possible level of glycation damage to tissues, and thus of diabetic complications, if continued for years.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for HbA1c has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HbA1c present is bound by the immobilized antibody. Anenzyme-linked monoclonal antibody specific for HbA1c is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of HbA1c bound in the initial step. The color development is stopped and the intensity of the color is measured.
