Velo™ Vitamin D Rapid Test (Whole Blood/Serum/Plasma)

• Velo^TM Vitamin D Rapid Test is an immunoassay based on the principle of sandwich During testing, the mixture migrates upward on the membrane chromatographically by capillary action. The membrane is pre-coated with 25 (OH) D antigens on the test line region (T). During testing, the specimen reacts with the particle coated with 25 (OH) D antigens. The mixture migrates upward on the membrane chromatographically by capillary action to react with 25 (OH) D antigens on the membrane and generates a colored line(s). The presence of this colored line in the test line region (T), indicates a positive result, while its absence indicates a negative result.
• An internal quality control is included in the test, in the form of a colored line appearing in the control line region (C), indicating that the test is functional, and proper and sufficient volume of specimen has been applied to enable migration through the test and control line, regardless of whether there is a test line or If the control line (C) does not appear within the testing time, test result is invalid, and the test should be repeated with a new test device.

Consider any materials of human origin as infectious and handle them using standard biosafety procedures. The test can be performed using whole blood (from venipuncture or fingerstick), serum or plasma specimens. Follow standard laboratory procedures to collect specimens.

For Whole Blood Collected by Fingerstick Using a Safety Sterile Blood Lancet (see illustrations below):

1. Tear off to open the Alcohol Prep Pad. Clean the area to be lanced with an alcohol swab.
2. Carefully twist off the protective cap until it is separated from the device.
3. Pace the lancet firmly against the puncture site to activate. Do not remove the device until an audible click is heard.
4. Squeeze the end of the fingertip, using a disposable dropper (30μL) provided, dip the tip end of the dropper into the blood specimen to collect fingerstick whole blood until reaching the 30μL specimen line, automatically.

NOTE:

• Use a new disposable dropper for each specimen in  order to avoid  cross contamination of specimens, which could  produce erroneous results.
• Discard the lancet or alcohol swab if package is pierced or The item may no longer be sterile; there is risk of infection if used.

For Whole Blood Collected by Venipuncture:

1. Using standard phlebotomy procedure, collect a venipuncture whole blood specimen using a blood collection tube with suitable anticoagulant (EDTA recommended).
2. It is recommended that specimens should be tested immediately. Do not leave the specimens at room temperature for prolonged periods. If the specimens are not tested immediately, they may be stored at 2°C-8°C.
3. It’s not recommended to test the whole blood specimens storing at 2°C-8°C for more than 2 days.

NOTE:

• Do not freeze a whole blood specimen, otherwise the red blood cell will break, which may cause hemolysis.
• Whole blood specimens should be stored in refrigeration (2-8°C) if not tested immediately. The specimens must be tested within 24 hours after collection.
• Anticoagulants including heparin, EDTA and sodium citrate do not affect the test results. Use of other anticoagulants have not been validated. Their use may affect the test result.

Plasma/Serum

1.  Collect blood specimen into collection tube containing EDTA, citrate or heparin for plasma or collection tube containing no anticoagulants for serum by venipuncture.
2. To make plasma specimen, centrifuge collected specimens and carefully withdraw the plasma into a new pre-labeled tube.
3. To make serum specimen, allow blood to clot, then centrifuge collected specimens and carefully withdraw the serum into a new pre-labeled tube.

Test the specimens as soon as possible after collecting. Store specimens at 2-8°C if not tested immediately. Specimens can be stored at 2-8°C for up to 3 days and should be frozen at -20°C for longer storage.

NOTE:

• Avoid multiple freeze-thaw cycles (no more than 3 times). Prior to testing, equilibrate frozen specimens to room temperature slowly and mix gently.
• Specimens containing visible particulate matter should be clarified by centrifugation before testing.
• Do not use samples demonstrating gross lipemia, gross hemolysis or turbidity so as to avoid interference on result interpretation.

Procedures

Before testing, open the package and equilibrate the test cassette, buffer, specimens and/or controls to room temperature, and shake the buffer gently before use. The most suitable temperature condition to perform the test is room temperature (15-30°C). If the test kit is stored at room temperature, it can be opened and used immediately.

1. Take out the test device from sealed foil pouch and place on a dry, clean and level surface.
2. Be sure to label the device with specimen’s ID number.
3. Uncap the buffer tube. Draw 20μL serum or plasma specimen, or 30μL whole blood specimen, with a laboratory transfer pipette or fingerstick dropper (30uL) as needed and add to the buffer tube. Screw on the cap and tighten securely to close the buffer tube. Shake the buffer tube vigorously to mix well the specimen with buffer. Then add three drops of mixed specimen to the specimen well (S) on the test cassette. See illustrations below.
4. Start the timer.
5. Wait for the colored line(s) to appear. Read test results at 15 minutes. Do not interpret the result after 20 minutes.

Internal Control: An internal quality control is included in the test, in the form of a colored line appearing in the control line region (C), indicating that the test is functional, and proper and sufficient volume of specimen has been applied to enable migration through the test and control line, regardless of whether there is a test line or not. If the control line (C) does not appear within the testing time, test result is invalid, and the test should be repeated with a new test device.
External Control: Control standards are not supplied with this kit; however, it is recommended that positive and negative controls be tested as a good laboratory practice to confirm the test procedure and to verify proper test performance.

• Positive: Two lines appear. One colored line should be in the control line region (C) and another apparent colored line(s) should be in the test line region (T).

NOTE: The intensity of the color in the test line region will vary depending on the concentration of Vitamin-D present in the specimen. Therefore, the presence of any test line, no matter how faint, within the designated observation time, indicates a positive result.

• Negative: One colored line appears in the control line region (C). No line appears in the test line region.
• Invalid: Control line fails to appear. Insufficient specimen volume or incorrect procedural techniques are the most likely reasons for control line failure. Review the procedure and repeat the test with a new test device. If the problem persists, stop using the test kit immediately and contact your local distributor.

• Clinical performance: Velo^TM Vitamin D Rapid Test been has correctly identified specimens of a performance panel and has been evaluated with a reference commercial enzyme-linked immunosorbent assay (ELISA) test using clinical specimens. Test results are presented in the table below.

Clinical performance compared to ELISA

Sensitivity (Positive Percent Agreement): 98.83% = 170/172 (95% CI: 95.86%~99.68%)

Specificity (Negative Percent Agreement): 99.25% = 268/270 (95% CI: 97.34%~99.80%)

Accuracy (Overall Percent Agreement): 99.09% = (170+268)/442 (95% CI: 97.70%~99.65%)

• Precision

Within-run and between-run precisions have been determined by testing 10 replicates on the same four specimens: a negative, a weak positive, a medium positive and a strong positive. The negative, weak positive, medium positive and strong positive specimens were correctly identified in all of the tests performed during each run.

• Cross-reactivity

No cross-reactivity was observed by testing the following positive specimens respectively: HAMA, HBsAg, HBsAb, HBeAg, HBeAb, HBcAb, HIV, HCV, H. pylori, MONO, CMV, Rubella and TOXO.

• Interference

The following potentially interfering substances were added to Vitamin D negative and positive specimens. Test results demonstrate that performance of VeloTM Vitamin D Rapid Test was not affected by the listed potentially interfering substances at the concentrations tested.

This test has been developed for testing human whole blood, serum, or plasma specimens only. The results of VeloTM Vitamin D Rapid Test should be evaluated with all clinical and laboratory data available. If test results do not agree with the clinical evaluation, additional tests should be performed. Other factors may interfere with VeloTM Vitamin D Rapid Test and may cause erroneous results. These include technical or procedural errors, as well as additional substances in blood specimens.

• The test kit should be stored either at room temperature or refrigerated (2°C-30°C), away from direct Do not freeze the kit or expose the kit to temperatures over 30°C.
• The shelf life of the kit is as indicated on the outer package (24 months from date of manufacture).
• This test kit is stable until the expiration date marked on the outer package and foil Ensure all test components are at room temperature (15-30°C) before use.
• Perform the test within one hour after taking out the test cassette from the foil pouch.