2.0 Indication: For In Vitro Diagnosis of colorectal, breast, lung and pancreas cancers
3.0 Package Size: One test per kit
4.0 Active Ingredients: 4.1 Coated Antibodies: 4.1.1 Control region: Goat anti-mouse (IgG) polyclonal antibody 4.1.2 Test region: Mouse monoclonal anti-CEA antibody A 4.2 Labeled Antibodies: Colloidal gold conjugate of monoclonal anti-CEA antibody B
5.0 Assay Protocol: 5.1 Assay Procedure: Read the package insert for WOHBC One-Step CEA test carefully before starting the assay. Allow serum samples to warm up to room temperature before testing. 5.1.1 Remove the test strip from its foil pouch and use it as soon as possible. 5.1.2 Immerse the test strip into the serum sample with the arrow end toward the serum for about 10 seconds. Do not immerse past the marker line. Take the strip out and lay it on a dry, clean and non-absorbing surface. 5.1.3 Wait 10 minutes and read results. It is important that the background is clear before the result is read. Results obtained after 30 minutes may be considered invalid.
5.2 Interpretation of Results: 5.2.1 Negative: Only one color band appears on the control region. No apparent band on the test region. 5.2.2 Positive: Distinct color bands appear on the control and test regions. Both test line and control line indicates a positive CEA result. Color intensity of the test bands may vary. 5.2.3 Invalid: No visible band at all or no colored line appears on the control region. Repeat test with a new test kit.
6.0 Cut off Value: 5 ng/mL CEA in serum sample
7.0 Storage: 2 to 30oC
8.0 Expiration Dating: 12 months from date of manufacture
9.0 Method of Manufacture: 9.1 Nitrocellulose Membrane Manufacture: 9.1.1 The purified monoclonal antibody, diluted in phosphate buffer saline, is coated on the test region. Simultaneously, goat anti-mouse IgG polyclonal antibody, diluted in phosphate buffer saline, is coated on the control region. 9.1.2 The coated membrane is dried for a minimum of 24 hours then sealed in an aluminum bag which contains silica gel desiccant. 9.2 Anti-CEA colloidal gold conjugate pad manufacture: 9.2.1 A buffer solution containing mouse Anti-CEA monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets. 9.2.2 The coated non-woven cloth is dried for minimum 24 hours then sealed in an aluminum bag which contains desiccant. 9.3 Test Device Assembly: 9.3.1 The coated membrane S1, the conjugate pad S4, and an absorbent pad is applied to an adhesive-coated backing. 9.3.2 A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, the assembled sheet of material is cut into strips. The test strips are then vacuum-dried for a minimum of 4 hours. 9.3.3 The assembled test strip is sealed in an aluminum pouch along with a desiccant packet.
10.0 Clinical Trials: To establish the sensitivity and specificity of WOHBC One-Step CEA test kit relative to other rates of qualitative serum tests, 203 samples from normal screening test and 115 samples from patients suspected of having colorectal, breast, lung and pancreas cancers were studied. The overall accuracy of WOHBC One-Step CEA test kits in this study was 95.28%, i.e., 303 out of 318 samples got the expected results when compared to the clinical follow up results.
11.0 Stability Studies: It is well known that immunochemicals are quite stable at room temperature if the materials are kept in dry condition. The chemicals used in the One-Step CEA device are either dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. Accelerated testing has been carried out in this study to establish the shelf life and to predict the expiration date of the test.
Three lots of One-Step CEA test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 42°C or room temperature. The test kits were removed from their storage and tested with positive and negative control panels. The results are shown in Table 2 and 3.
The data collected to date indicate that One-step CEA test kits are stable for at least 12 months at 2-30°C.
12.0 Quality Control: 12.1 Appearance: All materials are visually inspected before their use in manufacture. 12.2 Specificity: 10 test kits are randomly selected and tested with CEA<5ng/mL control panel. All test results must be negative. 12.3 Cut Off: 10 test kits are randomly selected and tested with a control panel in which contain 5ng/mL CEA. All test results must be positive. 12.4 Internal Control: A colored band must appear in the control region of the membrane with each tested kit, which indicating proper performance and reagent reactivity. 12.5 Reproducibility: Randomly selected test kits from different lots must give the same results when assaying the same sample.