Intended Use
The Quantitative Determination of Anti-H. Pylori Specific Antibodies of the IgG type in Human Serum or Plasma by Microplate Enzyme Immunoassay, Colorimetric.
Contents of Kit
A. Anti-H. Pylori Calibrators (Icons A-E)
Five (5) vials of references for anti-H. Pylori at levels of 0 (A), 10 (B), 25 (C), 50 (D), and 100 (E) U/mL of the IgG type. 1 mL/vial. Store at 2-8 °C. A preservative has been added.
B. Anti-H. Pylori Biotin Reagent (Icon)
One (1) vial containing biotinylated inactivated H. Pylori (IgG) in a buffering matrix. 13 mL/vial. A preservative has been added. Store at 2-8 °C.
C. Anti-H. Pylori Enzyme Reagent (Icon E)
One (1) vial containing anti-human IgG-horseradish peroxidase (HRP) conjugate in a buffering matrix. 13 mL/vial. A preservative has been added. Store at 2-8 °C.
D. Streptavidin Coated Plate (Icon)
One 96-well microplate coated with streptavidin and packaged in an aluminum bag with a drying agent. Store at 2-8 °C.
E. Serum Diluent
One (1) vial of containing buffer salts and a dye. 20 mL/vial. Store at 2-8 °C.
F. Wash Solution Concentrate (Icon)
One (1) vial containing a surfactant in buffered saline. 20 mL/vial. A preservative has been added. Store at 2-8 °C.
G. Substrate A (Icon SA)
One (1) vial containing tetramethylbenzidine (TMB) in buffer. 7 mL/vial. Store at 2-8 °C.
H. Substrate B (Icon SB)
One (1) vial containing hydrogen peroxide (H2O2) in buffer. 7 mL/vial. Store at 2-8 °C.
I. Stop Solution (Icon STOP)
One (1) vial containing a strong acid (1N HCl). 8 mL/vial. Store at 2-8 °C.
J. Product Instructions
Performance Characteristics
1. Precision
The within and between assay precision of the Anti-H. Pylori AccuBind ELISA Test System were determined by analyses on two different levels of pool control sera.
1.1 Precision Anti-H. Pylori - IgG
Within Assay Precision: Negative Pool - CV 5.6%, Positive Pool - CV 4.3%
Between Assay Precision: Negative Pool - CV 6.9%, Positive Pool - CV 5.0%
1.2 Precision Anti-H. Pylori - IgM
Within Assay Precision: Negative Pool - CV 7.4%, Positive Pool - CV 4.1%
Between Assay Precision: Negative Pool - CV 8.9%, Positive Pool - CV 7.5%
1.3 Precision Anti-H. Pylori - IgA
Within Assay Precision: Negative Pool - CV 8.5%, Positive Pool - CV 5.3%
Between Assay Precision: Negative Pool - CV 8.0%, Positive Pool - CV 7.6%
2 Sensitivity
The sensitivity (detection limit) was ascertained by determining the variability of the '0 U/mL' calibrator and using the 2 sigma (95% certainty) statistic to calculate the minimum dose:
Anti-H. Pylori IgG: 0.1424 U/mL
Anti-H. Pylori IgM: 0.065 U/mL
Anti-H. Pylori IgA: 0.304 U/mL
3 Accuracy
The Anti-H. Pylori AccuBind ELISA Test System was compared with a reference ELISA method. Biological Specimens from varying concentrations were assayed.
Precision
Anti-H. Pylori IgG:
Within Assay: Negative CV 5.6%, Positive CV 4.3%
Between Assay: Negative CV 6.9%, Positive CV 5.0%
Detection Range
0 - 100 U/mL
Detection Limit
IgG: 0.1424 U/mL
IgM: 0.065 U/mL
IgA: 0.304 U/mL
Sensitivity
The sensitivity (detection limit) was ascertained by determining the variability of the '0 U/mL' calibrator and using the 2 sigma (95% certainty) statistic to calculate the minimum dose:
Anti-H. Pylori IgG: 0.1424 U/mL
Anti-H. Pylori IgM: 0.065 U/mL
Anti-H. Pylori IgA: 0.304 U/mL
General Description
Helicobacter pylori has been shown to be the bacillus that was observed by Warren and Marshall in close contact with gastric epithelium in biopsy studies of patients suffering from chronic gastritis. Although the source of H. Pylori infection is not known, the evidence is quite convincing that the bacillus can cause acute gastritis and may lead to chronic gastritis. H. Pylori was present in ninety-one (91) percent of patients with chronic superficial gastritis. H. Pylori was present in ninety (90) percent of duodenal ulcer and seventy (70) percent of gastric ulcer patients. The use of serological testing to ascertain the immunologically produced antibody caused by H. pylori infection has been suggested as the method of choice to screen large populations.
Standard Curve
Cal A (0 U/mL): 0.044 Abs
Cal B (10 U/mL): 0.406 Abs
Cal C (25 U/mL): 0.782 Abs
Cal D (50 U/mL): 1.293 Abs
Cal E (100 U/mL): 2.354 Abs
Reconstitution And Storage
Store all reagents at 2-8 °C for stability until the expiration date printed on the kit. Avoid extended exposure to heat and light. Opened reagents are stable for sixty (60) days when stored at 2-8 °C. Kit and component stability are identified on the label.
Note 1: Do not use reagents beyond the kit expiration date.
Note 2: Avoid extended exposure to heat and light. Opened reagents are stable for sixty (60) days when stored at 2-8 °C.
Note 3: Above reagents are for a single 96-well microplate.
Citations
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