Kit for screening and checking glucose-6-phosphate dehydrogenase (G6PD) deficiency of neonates and preparation method for kit

A hexaphosphate dehydrogenase and glucose technology, which is applied to a kit for screening neonatal glucose hexaphosphate dehydrogenase deficiency and its preparation field, can solve the problems of high false positive rate and low accuracy rate of blood samples, and achieves The effect of reducing false positive rate, high accuracy and low cost

Active Publication Date: 2012-06-27
GUANGZHOU FENGHUA BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the fluorescence analysis method has a high false positive rate and a low accuracy rate for anemia, hemolysis, coagulation and old blood samples.

Method used

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  • Kit for screening and checking glucose-6-phosphate dehydrogenase (G6PD) deficiency of neonates and preparation method for kit
  • Kit for screening and checking glucose-6-phosphate dehydrogenase (G6PD) deficiency of neonates and preparation method for kit
  • Kit for screening and checking glucose-6-phosphate dehydrogenase (G6PD) deficiency of neonates and preparation method for kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The components of the simultaneous detection G6PD assay kit (fluorescence ratio method) of the present invention include:

[0051] (1) G6P substrate reagent dry powder: made of G-6-P and NADP and freeze-dried;

[0052] (2) 6PG substrate reagent dry powder: made of 6-PG and NADP and freeze-dried;

[0053] (3) Copper reagent: made of copper sulfate, potassium sodium tartrate and sodium carbonate;

[0054] (4) G6PD filter paper dry blood spot quality control, C1 positive, C2 negative: use anticoagulated sheep whole blood red blood cells to prepare filter paper dry blood spot;

[0055] (5) Reagent for substrate reconstitution: made with MgCl2, Gly-Gly (diglycine) and Tris-Hcl.

[0056] (6) Disc quality control: citrate anticoagulated sheep whole blood in a low-speed centrifuge at 3000rmp, and centrifuge for 10-15min to separate plasma and red blood cells. Adjust the hematocrit according to the ratio (red blood cell: plasma = 1.2: 1.0); add G6PD and 6PGD pure raw material...

Embodiment 2

[0068] Embodiment 2 The usage method of kit of the present invention

[0069] The specific operation of the neonatal G6PD screening kit (fluorescence ratio method) prepared in Example 1 above is as follows:

[0070] Reagent preparation:

[0071] Substrate redissolution: G6P substrate reagent dry powder, 6PG substrate reagent dry powder respectively add 16ml substrate redissolution reagent, mix well, let stand for 20 minutes;

[0072] Cooling of the copper reagent: the reagent must be cooled in a refrigerator at 4°C before use.

[0073] Check operation:

[0074] Adding samples: Punch out two samples with a diameter of about 3mm (1 / 8 inch) from the sample filter paper dry blood sheet with punching forceps (the paper sheet must be soaked with blood), and add the blanks of G6PD and 6PGD respectively in turn In the microwell, each hole corresponds to one piece. Control the consistency of punching to make the paper as consistent as possible.

[0075] Add substrate: add 150 μl o...

Embodiment 3

[0080] Example 3 Analysis performance evaluation index of the kit of the present invention

[0081] The performance evaluation indexes of the newborn screening assay G6PD kit (fluorescence ratio method) prepared in the above Example 1 are as follows:

[0082] Sensitivity: determine 1 set of positive reference products established by the company's laboratory. Positive reference products are samples from patients with G6PD deficiency determined by clinical testing, consisting of 100 samples.

[0083] Conformity rate of positive reference products (%) = measured number of positive reference products / 10*100%.

[0084] Table 2 Sensitivity test of kit of the present invention

[0085] Positive (ratio ≤ 1.0) Negative (ratio > 1.0) Test results 100 0

[0086] Continuing through the above table, the positive coincidence rate is 100%.

[0087] specificity:

[0088] Determination of conformity rate of negative reference products: 1 set of negative reference...

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Abstract

The invention discloses a kit for screening and checking glucose-6-phosphate dehydrogenase (G6PD) deficiency of neonates and a preparation method for the kit. The kit for screening and checking the glucose-6-phosphate dehydrogenase (G6PD) deficiency of the neonates consists of substrate reagent dry powder, a substrate redissolved reagent, a copper reagent, a reaction board and filter paper dried blood spot quality control material. The detection result of the kit is high in accuracy and sensitivity, and the kit has the advantages of high stability, economy, simplicity, high efficiency and thelike; when the kit is applied to screening and checking for the neonates, the relevance ratio of heterozygote is improved; and the judgment accuracy is improved.

Description

technical field [0001] The invention relates to the field of detection kits, in particular to a kit for screening neonatal glucose-6-phosphate dehydrogenase deficiency and a preparation method thereof. Background technique [0002] Glucose-6-phosphate dehydrogenase deficiency is a group of heterogeneous diseases caused by a significant deficiency of erythrocyte glucose-6-phosphate dehydrogenase (G6PD), commonly known as "faba bean disease", and is the most common inherited enzyme deficiency disease. Glucose-6-phosphate dehydrogenase deficiency (Glucose-6-phosphate dehydrogenase G6PD) is one of the important housekeeping enzymes of almost all high and low organisms, with a molecular weight of 59kDa, and its active form consists of 2 or 4 subunits, each Each subunit contains 515 amino acids. The enzyme catalyzes the rate-limiting step of the hexose phosphate bypass pathway in the process of cellular energy metabolism, and in the process reduces NADP (coenzyme Ⅱ) to NADPH, whi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/52
Inventor 吴道贫冯健明汪勤沈健何海荣孙勇赵可辉王云爱
Owner GUANGZHOU FENGHUA BIOENG
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