Glucose-6-Phosphate Dehydrogenase (G6PD) Assay Kit – Bulk Reagents

US$167.00

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Selling unit: per 100ml

Product Name

Common name: Glucose-6-phosphate dehydrogenase (G6PD) assay kit (rate method)

English name: Glucose-6-phosphate Dehydrogenase Reagent Kit (Kinetic Method)

Reagent Ratio

3:1,4:1 can be used.

Intended Use

Used to determine the activity of glucose-6-phosphate dehydrogenase (G-6-PD) in human whole blood.

G-6-PD deficiency is a common hereditary enzyme deficiency, commonly found in the Yangtze River basin and provinces south of China, with a higher incidence in males than females due to the X-linked incomplete dominant inheritance of the gene. It is also reduced in acute leukemia, diabetes mellitus, neonatal jaundice, etc.; it is increased in hepatocellular diseases and drug hemolysis. Screening for this indicator in people with high prevalence can effectively prevent the occurrence of hemolysis, and premarital physical examination and prenatal examination are important for eugenics and effective prevention of neonatal jaundice.

Test Principle

G-6-PD catalyzes the formation of glucose 6-phosphate and NADPH from glucose-6-phosphate and NADP, causing an increase in absorbance at 340 nm. The activity of G-6-PD can be measured by monitoring the rate of increase in absorbance at 340 nm.

G6PD

Glucose-6-phosphate + NADP ──→ 6-phosphoglucuronide + NADPH

Main Components

Reagent components included in the product.

Glucose-6-Phosphate Dehydrogenase (G6PD) Assay Kit

*Components are not interchangeable in kits with different lot numbers.

*Reagent components not included in the product, but necessary for the test: outsourced normal and abnormal QCs and calibrators.

Storage Conditions And Expiration Date

  1. The unopened reagent can be stored at 2 – 8℃ away from light and can be stabilized until the expiration date, which is one year.
  2. Take care to avoid contamination of reagents that have been opened.
  3. After opening the bottle, the reagent should be stored at 2~8℃ for 2 weeks to keep it stable from light, and the reagent should not be frozen.
  4. Use water as blank solution, reagent blank absorbance absorbance greater than 1.0 is prohibited.

Applicable Instruments

Hitachi 7180/7600; Olympus AU680/2700; Toshiba TBA120; Myriad BS2000M/480; Siemens ADVIA 1800/2400 series automatic biochemical analyzers.

Sample Request

1.Sample type: whole blood specimen processing as well as preservation: EDTA-K anticoagulated whole blood, (2-8)℃ can be stored for 1 week.

  1. 2.sample collection: take EDTA-K anticoagulated whole blood, centrifuge for 5 minutes at 3000 rpm, remove the supernatant, aspirate 20ul of lower pressure red blood cells into 1ml of deionized water (diluted 51 times), mix thoroughly and wait for the red blood cells to dissolve completely (15-30 minutes) for determination.

3.Sample interference: samples with interference to the reaction absorbance, including samples of hemolysis and lipidemia may affect the test results, in the above case, it is recommended to re-collect.

Test Method

  1. Reagent preparation: liquid reagents are ready to use out of the bottle.
  2. test conditions: (different test instruments can be requested according to the different parameters on the machine)

Glucose-6-Phosphate Dehydrogenase (G6PD) Assay Kit

Operation steps.

Glucose-6-Phosphate Dehydrogenase (G6PD) Assay Kit

Reagents and sample volumes can be increased or decreased in proportion to the requirements of different biochemical analyzers.

  1. Calibration procedure.

Using the theoretical factor, deionized water was used as the blank calibration. Theoretical factor (F-value): 4019

Requirements for calibration and calibration frequency: Under normal circumstances, the measurement should be calibrated at least once every 2 weeks.

  1. QC control procedures.

It is recommended that each laboratory establish its own quality control system and select appropriate quality control products for quality control. The measured values of QC products should be within the specified range. If outside the specified range, it is necessary to take appropriate measures or contact the manufacturer.

  1. Calculation.

ΔA/min×Vt×1000

Lysate blood G6PD activity (U/L) = ——————————– = ΔA/min×F1*

e×Vs×d

G6PD activity (U/L) in pressurized erythrocytes = lysate blood G6PD activity × 51 = F2* × ΔA/min (F2*= F1* × 51 = 204970)

Positive Judgment Value Or Reference Interval

Normal reference range for adult G6PD in compressed red blood cells.

* Using EDTA anticoagulated blood, greater than 1300 U/L; samples with levels of 600-1300 U/L are suspicious specimens and can be tested for G6PD activity per gram of hemoglobin according to the calculation in Appendix 2), or for further examination.

* Using heparin anticoagulated blood, greater than 1100 U/L. Samples with levels of 500-1100 U/L are suspicious specimens and can be tested for G6PD activity per gram of hemoglobin according to the calculation in Appendix 2) or for further examination.

* G6PD activity per gram of hemoglobin, normal value range: >3.8 U/gHb.

Interpretation Of Test Results

If the results exceed the linear range, dilute the specimen 1:1 with physiological saline and multiply the measurement by 2.

Professional staff is responsible for reviewing the test results. Test results can be affected by the age, gender, and weight of the person being tested. Normally, the results are considered normal if they are within the reference range; if they are within the critical region, they should be re-measured for confirmation; if they are significantly outside the reference range or still outside the reference range after confirmation testing, the target level in the serum is considered abnormal. If the test results do not match or even contradict the clinical situation, the reasons should be analyzed and found.

Limitations Of The Test Method

The specificity of this method is limited and it can only be used as a screening test, not to confirm the diagnosis. If a suspicious specimen is encountered, the diagnosis should be confirmed by definitive methods. The activity of the substance to be measured is relatively stable in red blood cells, but after hemolysis treatment, the activity is lost quickly, and can be reduced by 30% after 1 hour, so the sample should be measured as soon as possible after processing.

Product Performance Index

Appearance: R1: colorless and clarified liquid; R2: colorless and clarified liquid. QC: colorless or light yellow clarified liquid.

Blank absorbance: A (blank) <0.3 ABS ( 1 cm; 340 nm; 37 °C).

Blank absorbance change rate: (ΔA/min) should be ≤ 0.003 ABS/min ( 1cm; 340nm; 37℃).

Analytical sensitivity: absorbance change ≥ 0.008 at a concentration of 50 U/L.

Linearity of reagent determination: linear correlation coefficient r2 ≥0.995 in the linear range of (0 – 1500) U/L. Relative deviation ≤10% in the range of (401 – 1500) U/L; absolute deviation ≤40 U/L in the determination of (0 – 400) U/L. (Note: Since the sample is diluted 51 times, the reagent can be monitored in the range of (0 – 1500×51)) U/L)

Precision: The precision CV of the kit test items is < 5 %.

Inter-batch variation: The relative deviation of the measurement results between different batches should be < 6 %.

Accuracy: The measured value is within the quality control target value when testing the quality control product of Beijing Lidman Biochemical Co.

Caution

  1. This product is for in vitro diagnosis only.
  2. Avoid contamination when using the reagent, the container used must be clean, and please take necessary precautions, do not swallow, and avoid contact with skin and mucous membrane.
  3. Please dispose of the measured samples and waste liquids in accordance with the relevant national and local laws and regulations.
  4. Do not mix R1 and R2 into a single reagent.
  5. When changing the reagent lot number, please recalibrate.

Reference

Young, D.S., et al: Clin. Chem. 20:10 (1975).

Remaly AT, Wilding P. Macro enzymes: Biochemical Characterization, Clinical Significance, and Laboratory Detection. Clin Chem 1989;35: 2261 -2270.

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