25?hydroxy Vitamin D Test Kit (Homogeneous Chemiluminescence Immunoassay)

【Product Name】

25?hydroxy Vitamin D Test Kit (Homogeneous Chemiluminescence Immunoassay)

【Packing Specification】

25 tests/ box.

【Intended Use】

This assay is intended for the quantitative determination of total 25-hydroxyvitamin D in human serum and plasma. This assay is to be used as an aid in the assessment of vitamin D sufficiency.

Vitamin D is a fat-soluble steroid hormone precursor that is mainly produced in the skin by exposure to sunlight. Vitamin D is biologically inert and must undergo two successive hydroxylations in the liver and kidney to become the biologically active 1,25-dihydroxyvitamin D.

Most of the 25-hydroxyvitamin D, measurable in serum, is 25-hydroxyvitamin D3 whereas 25-hydroxyvitamin D2 reaches measurable levels only in patients taking vitamin D2 supplements. Vitamin D2 is considered to be less effective.

The most abundant product of 25-hydroxyvitamin D catabolism by 24-hydroxylase (CYP24A1) is 24,25-dihydroxyvitamin D.6 It accounts for 2-20 % of the total circulating 25-hydroxyvitamin D, has a half-life of approximately 7 days and is present in serum at concentrations of up to approximately 10 nmol/L.

Vitamin D is essential for bone health. In children, severe deficiency leads to bone-malformation, known as rickets. Milder degrees of insufficiency are believed to cause reduced efficiency in the utilization of dietary calcium.

Vitamin D deficiency causes muscle weakness; in elderly, the risk of falling has been attributed to the effect of vitamin D on muscle function.

Vitamin D deficiency is a common cause of secondary hyperparathyroidism. Elevations of parathyroid hormone levels, especially in elderly vitamin D deficient adults can result in osteomalacia, increased bone turnover, reduced bone mass and risk of bone fractures. Low 25-hydroxyvitamin D concentrations are also associated with lower bone mineral density. In conjunction with other clinical data, the results may be used as an aid in the assessment of bone metabolism.

So far, vitamin D has been shown to affect expression of over 200 different genes. Insufficiency has been linked to diabetes, different forms of cancer, cardiovascular disease, autoimmune diseases and innate immunity.

【Principle】

Competition principle.

The 25?hydroxy Vitamin D?test kit employs a wash-free and homogeneous strategy based on proximity hybridization-regulated CRET(Chemiluminescence resonance energy transfer). The AE demonstrated strong chemiluminescence (CL) in the presence of H2O2. Graphene oxide (GO) as an excellent quencher was used to produce the “Signal off” mode that little CL emission was observed through CRET between GO and the AE-labelled DNA3. Once the antigen was introduced, the target-induced proximity hybridization occurred to form a proximate complex, which inhibited the CRET by preventing GO from absorbing AE-labelled DNA3. The CL intensity decreased with the increasing 25OH Vitamin D?concentration.

【Main Components】

Lyophilization sphere reagents: DNA1-labeled 25-OH VD antibody (recombinant human IgG) (20ng/person), DNA2-labeled 25-OH VD-BSA (10ng/person), DNA3-labeled acridine ester (AE), graphene oxide (GO) conjugated antioxidant (AOD), preservative (P300) (0.1%).

Dissociation solution: composed of phosphate buffer (10 mmol/L), preservative (0.1%), etc.

Calibration card: containing standard curve for product calibration.

※The components in the kits of different batches are not interchangeable.

2?Reaction cups

2?Buffer solution

2?Substrate solution

2?Pipette

2?Disposable tip

【Storage Conditions and Expiration Date】

Stable for 18 months when stored at 2-8℃?away from light. Reagents are used immediately upon opening.

【Applicable Instruments】

Chemiluminescence immunoassay analyzer (HSCL-5000)?produced by Nanjing Poclight Biotechnology Co. Ltd.

【Sample Requirements】

1. The serum is recommended to be collected by ordinary serum collection tubes or tubes with procoagulant. If it cannot be used immediately, it is recommended to be stored at 2-8℃ and the test should be completed within 24 hours. If the test cannot be performed within 24 hours, the sample needs to be frozen at -20℃ for 1 month.

2. Samples containing sediment or flocculent material should be centrifuged (4000rpm, 10min) and then tested.

3. Samples must be returned to room temperature (15-30℃) before testing. Frozen samples should be completely thawed, rewarmed and mixed well before use. Do not repeatedly freeze and thaw.

4. Heat inactivated samples, hemolysis and other abnormal samples should be discarded.

【Test Method】

The test method is homogeneous chemiluminescence immuoassay

1. Preparation before testing:

1.1 Please read this manual and the instrument manual carefully before use.

1.2 Check whether the instrument can work normally, whether the reaction cup or sample tube, substrate solution and buffer are prepared adequately, and prepare other related consumables.

2. Test procedure

2.1 Turn on the chemiluminescence?immunoassay analyzer.

2.2 Check whether the calibration card is consistent with the lot number of the kit and scan the calibration card.

2.3 Absorb 100μL of the sample to be tested and 200μL of the dissociation solution into the reaction cup and mix well;?put into the instrument;?start incubation,?measurement,?processing data;?and derive the test results.

2.4 Total assay time is 10 minutes.

3.?Result analysis.

The measured chemiluminescence intensity is converted from the standard curve to obtain the concentration of 25-OH VD in the unknown sample, which can be read at the ng/mL level.

4. Calibration

Each kit batch contains its specific calibration information, which is stored on a calibration card.

【Reference Interval】

The following reference ranges were determined by statistical analysis of the 95% distribution range of 25-OH VD levels in 200 healthy human samples.

Normal reference value: ≥30ng/mL

Clinical diagnostic threshold (Cut-off value): 25-OH VD ≤ 30 ng/mL predicts vitamin D deficiency or insufficiency

Note: Due to differences in geography, ethnicity, gender and age, it is recommended that each laboratory establish its own reference range.

【Interpretation of Test Results】

1. The test results are for clinical reference only and should be considered in conjunction with clinical symptoms, medical history, other laboratory tests and other circumstances, and are not to be used as a basis for clinical diagnosis.

2. If the test result is abnormal or differs from the clinical diagnosis, verification is recommended.

3. The operation must be done in strict accordance with the operation procedures and carefully in order to get the correct results. Any modification to the operating procedures may affect the accuracy of the test results.

4. Bacterial contamination of the sample or repeated freeze-thawing may also affect the results.

【Product Performance Index】

1. Appearance: complete components, lyophilized balls in complete form.

2. Accuracy: relative deviation not more than 8%.

3. Blanking limit: not more than 2ng/mL; detection limit: 3 ng/mL

4. Linearity: 3-100ng/mL, in this linear range, the linear correlation coefficient r should be not less than 0.990.

5. Repeatability: CV≤5%.

6. Inter-batch variation: CV≤8%.

【Caution】

1. This product is a single-use in vitro diagnostic reagent, please do not reuse it, and do not use the expired product.

2. The reagent can only be used on the supporting instrument analysis system.

3. The test procedure must be followed as indicated, improper operation will lead to wrong results.

4. Components of kits with different lot numbers cannot be mixed.

5. The calibration card used for calibration should not be exchanged with kits of different lot numbers.

6. Do not swallow the reagents or contact with skin, eyes or mucous membranes as the reagents contain preservatives; once contacted, rinse the contaminated area with water.

7. When using the instrument for testing, each sample and the corresponding reagents should be replaced with a new disposable tip to avoid cross-contamination, which may affect the accurate determination of 25-OH VD concentration in the sample, and all samples should be aspirated directly to the bottom of the sample tube.

8. The use of fresh samples is recommended. All samples from patients should be treated as potential sources of infection and all waste generated during the test must be disposed of in accordance with local regulations.

9. If the reagent has exceeded its expiration date, it should not be used again.

10. This kit contains animal-derived components that are potentially infectious.

【Production Date and Expiration Date】

See labels for details

【Basic Information】

Registrant/ Producer Name：Nanjing Poclight Biotechnology Co., Ltd

Address：South Side, 2nd?Floor, Building A02, Biopharmaceutical Valley Accelerator (Phase III), No. 142 Huakang Road, Jiangbei New District, Nanjing City, Jiangsu Province, P.R. China.

After-sales service unit：Nanjing Poclight Biotechnology Co., Ltd

Postal Code：210000

Website：https://www.puguangbio.com/