Interleukin?6 (IL-6) Test Kit (Homogeneous Chemiluminescence Immunoassay)

【Product Name】

Interleukin?6 (IL-6) Test Kit (Homogeneous Chemiluminescence Immunoassay)

【Packing Specification】

25 servings/ box.

【Intended Use】

Immunoassay for the in vitro quantitative determination of Interleukin-6 (IL-6) in serum and plasma. This assay can be used to aid in the management of critically ill patients as early indicator for acute inflammation.

Interleukin-6 (IL-6) is a pleiotropic cytokine with a wide range of functions. It was first described as interferon-β2, plasmacytoma growth factor, and hepatocyte stimulating factor. Later on it was described as human B-cell-stimulating factor 2 (BSF2). In 1988 it was proposed to name it IL-6 as further studies had demonstrated that the protein shows activities not only on B-cells but also on T-cells, hematopoietic stem cells, hepatocytes and brain cells. IL-6 is produced from a single gene encoding a product of 212 amino acids, which is cleaved at the N-terminus to produce a 184 amino acid peptide with a molecular weight between 22-27 kDa. In 1989 it was reported that also immunoreactive complexes in the range of 60-70 kDa were detected in human body fluids in patients with acute bacterial infections.IL-6 production is rapidly induced in the course of acute inflammatory reactions associated with injury, trauma, stress, infection, brain death, neoplasia, and other situations.

IL-6 concentrations in trauma patients may predict later complications from additional surgical stress or indicate missed injuries or complications.

Sequential measurements of IL-6 in serum or plasma of patients admitted to the ICU (intensive care unit) showed to be useful in evaluating the severity of SIRS (systemic inflammatory response syndrome), sepsis and septic shock and to predict the outcome of these patients.6,7,J8 IL-6 is also useful as an early alarm marker for the detection of neonatal sepsis.IL-6 also plays a role in chronic inflammation e.g. rheumatoid arthritis.

【Principle】

Sandwich principle.

The Interleukin?6 (IL-6)?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 increased with the increasing IL-6?concentration.

【Main Components】

Lyophilized sphere reagents: DNA1-labeled IL-6 1 antibody (mouse) (≤1.0ug/mL), DNA2-labeled IL-6 2 antibody (mouse) (≤1.0ug/mL), DNA3-labeled acridine ester (AE), graphene oxide (GO) conjugated antioxidant (AOD), preservative (P300) (0.1%).

Calibration card: containing standard curve for product calibration.

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

2?Reaction cups

2?Compound 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. Plasma is recommended to be anticoagulated with heparin or EDTA. If it cannot be used immediately, it is recommended to be stored at 2-8℃ to complete the test within 24 hours. If the test cannot be performed within 24 hours, the sample should be frozen at -20℃, valid for 1 month.

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

4. 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.

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

【Test Method】

The test method is homogeneous chemiluminescence immuoassay.

1. Preparation before the test.

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

1.2 Check whether the instrument can work properly, whether the reaction cup or sample tube, substrate solution and buffer preparation are sufficient, and prepare other related consumables.

2. Test procedure

2.1 Turn on the chemiluminescence?immunoassay analyzer.

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

2.3 Mix 50μL of the sample to be tested into the reaction cup, put it into the instrument and start incubation, measurement, data processing and test results.

2.4 The total detection time is 5 minutes.

3.Result analysis.

The measured chemiluminescence intensity is converted from the standard curve to obtain the concentration of IL-6 in the unknown sample, which can be read at the pg/mL level.

4. Calibration

Each batch of the kit 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% confidence interval distribution range of IL-6 levels in 200 healthy human samples according to normal distribution.

Normal reference value: 7 pg/mL

The measured IL-6 levels will vary depending on the region, individual differences and the assay method used. Therefore, it is recommended that each laboratory should establish its own reference range for its own characteristic population.

【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 should not be used as the sole 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.

5. Values below the lower limit of detection are reported as <1.5 pg/mL, and values above this measurement range are reported as >5000 pg/mL.

6. The amount of IL-6 in the sample is outside the test range and should be pre-diluted with saline before testing. The maximum dilution is 10 times, and the upper limit of the assay after dilution is 50,000 pg/mL.