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Ostase® BAP ELISA

INTENDED USE

The Ostase BAP Immunoenzymetric Assay is an in vitro device indicated for the quantitative measurement of bone-specific alkaline phosphatase (BAP), an indicator of osteoblastic activity, in human serum. This device is intended to be used as an aid in the management of postmenopausal osteoporosis and Paget’s disease.

SUMMARY AND EXPLANATION

Bone is a dynamic tissue in which bone formation and bone removal (also referred to as resorption) continue throughout life in a process called remodeling. The remodeling process is a function of complex interactions between two types of bone cells: osteoblasts for the formation of bone, and osteoclasts for the resorption of bone. Bone formation and resorption are interdependent processes that are, under normal circumstances, tightly coupled. This coupled relationship is integral to maintaining the biochemical competence of the skeleton, thereby preserving the organization of bone structure, form, and strength.

Serum levels of BAP are believed to reflect the metabolic status of osteoblasts. An accurate assessment of bone metabolism is critical for determining the severity of metabolic bone disease and responses to therapy. Measurement of serum levels of BAP has been shown to be useful in evaluating patients with Paget’s disease, osteomalacia, primary hyperparathyroidism, renal osteodystrophy, osteoporosis and metastases to bone. Total alkaline phosphatase determinations have been the accepted method for the diagnosis and monitoring of patients with Paget’s disease.

Paget’s disease of bone is a common skeletal disorder in which there is a focal proliferation of the normal cellular components of bone. Paget’s disease is more prevalent than once thought with the incidence rate in certain populations at 3%-4% in middle-aged patients and 10%-15% in the elderly . This disease does not affect young individuals. The majority of patients with Paget’s disease have no symptoms and often go undiagnosed unless an abnormal X-ray or serum alkaline phosphatase level is found in the course of a medical evaluation for unrelated reasons. The most common complaints in symptomatic patients are pain and deformity.

The risk of osteoporosis, another bone remodeling disorder, depends in part upon skeletal development,the attainment of peak bone mass, and in later life, the amount of bone lost. In healthy children,bone formation is favored over bone resorption, which results in bone development and normal skeletal growth. In healthy young adults, bone formation and bone resorption are balanced, resulting in no net increase or decrease in skeletal mass. With advancing age, men and women experience an imbalance in bone remodeling in which resorption is slightly greater than formation, resulting in a continuous net loss of bone mass with time. If this imbalance persists, bone mass may decline until the skeleton is insufficient to withstand normal mechanical stresses, and it becomes abnormally susceptible to fractures. The excessive loss of bone mass with an increased susceptibility to fractures is a disorder known as osteoporosis.

The most common form of osteoporosis occurs in postmenopausal women and is the result of estrogen

deficiency. Rapid bone loss accompanies the decline of estrogen levels at the onset of menopause or as a result of surgical removal of the ovaries. Rapid bone loss occurs as a result of the combined effects of an imbalance in bone remodeling and an increase in bone turnover. In the United States, osteoporosis affects some 25 million postmenopausal women and is the cause of approximately 1.5 million fractures annually, including approximately 500,000 vertebral crush fractures, 250,000 hip fractures, and 200,000 distal radius fractures.

Hormone replacement therapy is currently the most widely prescribed therapy for the prevention of osteoporotic fractures in postmenopausal women. However many women cannot, or will not,avail themselves of hormone replacement therapy because of the potential for the increased risk of cancer and the resumption of menstrual bleeding. For this reason, other compounds such as bisphosphonates,a standard treatment for Paget’s disease of bone, have been developed to treat osteoporosis. The antiresorptive properties of bisphosphonates decrease bone remodeling and, consequently, decrease the overall loss of bone.

Biochemical markers are useful in monitoring metabolic bone disease. Urinary hydroxyproline and total serum alkaline phosphatase have been used for monitoring the treatment of Paget’s disease.Osteoporosis, however, represents a more subtle modification of the bone remodeling process; therefore,more specific and sensitive markers are needed.

The Ostase BAP assay is an in vitro device for the quantitative measurement of bone-specific alkaline phosphatase (BAP) in human serum. Changes in BAP have been shown to be useful in patients undergoing therapy for metabolic bone disorders.

PRINCIPLES OF THE PROCEDURE

The Ostase BAP assay is a solid phase, monoclonal antibody immunoenzymetric assay. Samples containing BAP are reacted with a solution containing a biotin-labeled, BAP-specific monoclonal antibody.The reaction takes place in plastic well strips (solid phase) coated with streptavidin and enclosed in a plastic frame. Following the formation of a solid phase/capture antibody/BAP complex, the microplate is washed to remove unbound BAP and is then incubated with an enzyme substrate. The amount of substrate turnover is determined colorimetrically by measuring the absorbance of the quenched reaction at 405 nm in a microplate reader. The absorbance is proportional to the concentration of BAP present in the test sample. The calculation of BAP concentration in the sample is based on concurrent testing of BAP calibrators and Zero Calibrator/Diluent.

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