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Novel platelet cross-match kit

A technology of platelets and kits, which is applied in the field of kits and new platelet cross-matching kits, can solve the problems of low sensitivity and high price, and achieve the effects of reducing detection costs, strong adsorption capacity, and good clinical application value

Active Publication Date: 2019-10-08
广州血液中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The innovation of the present invention is to use the high adhesion of polyethyleneimine to coat it on the bottom of the microplate hole, innovatively use its adsorption and the flocculation of polymerized cations to produce high-strength adsorption on platelets, and use high-sensitivity secondary The one-step indicator system replaces the one-step indicator system, which can effectively solve the problems of low sensitivity and high price in the current clinical platelet cross-match method, and provide a cost-effective platelet cross-match kit for clinical front-line detection work

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The preparation of embodiment 1 platelet matching kit

[0026] Preparation of U-shaped reaction microplate

[0027] Prepare 0.1% polyethyleneimine solution with 0.05M, pH9.6 carbonate buffer solution, add 50uL / well into the wells of a 96-well U-shaped (12×8) reaction plate, and place it in a refrigerator at 4°C overnight. Take it out the next day, wash it with 1×PBS buffer solution for 3 times, shake off the washing solution, blot the remaining liquid in the hole with absorbent paper for the last time, pack the reaction plate in an aluminum foil bag, heat and seal it, and place it in a 4°C refrigerator Save for later.

[0028] platelet acquisition

[0029] Platelets can be directly prepared from machine-collected platelets or platelet-rich plasma. When using platelet-rich plasma to prepare platelets, take several milliliters of freshly collected EDTA anticoagulated whole blood, centrifuge at 1500rpm for 10 minutes, and absorb the upper 2 / 3 platelet-rich plasma. Afte...

Embodiment 2

[0041] Embodiment 2 platelet cross-matching type

[0042] Take the kit out of the refrigerator to equilibrate to room temperature, and extract the strips according to the number of tests.

[0043] Mark the extracted microplate strips, such as experimental wells, negative control wells, positive control wells, etc.

[0044] Add 100×109 / ml platelet suspension from different blood donors to the 6 reaction wells from top to bottom, 50uL per well, tap the side of the strip for mixing.

[0045] Put the reaction strips into a flat-bed centrifuge, centrifuge at 50g for 5 minutes, take out the strips, shake them upside down to remove the unbound platelet suspension, wash with 1×PBS for 3 times, and shake them upside down lightly each time Liquid, after the last light shake dry, buckle the reaction plank upside down on the absorbent paper, blot the residual liquid in the hole.

[0046] Add low-ionic media to the reaction wells, 100uL / well, then add patient serum or plasma to the exper...

Embodiment 3

[0051] Embodiment 3 kit stability test

[0052] The U-shaped reaction microplate prepared in Example 1 was stored in a refrigerator at 2-8°C, and samples were taken at 2 months, 4 months, 6 months, 8 months, 10 months, and 12 months of the storage period. Stability study; in addition, samples from patients with platelet antibodies were collected and stored in a -20°C refrigerator after aliquoting. Take 1 tube out of the refrigerator for each test and use it after thawing. As a result, it was found that the U-shaped reaction microplate prepared by the present invention had no difference in the use performance of the cross-matching type within 12 months of storage, reflecting better stability. The results are shown in image 3 .

[0053] 1. Take out the kit from the refrigerator and equilibrate to room temperature, and extract the strips according to the number of tests.

[0054] Mark the extracted microplate strips, such as experimental wells, negative control wells, positiv...

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Abstract

The invention discloses a novel platelet cross-match kit, which comprises a U-shaped reaction microplate, a low-ion medium, an indication system, a negative control serum, and a positive control serum, wherein the inner surface of the U-shaped reaction microplate is coated with polyethyleneimine, and by using the high adsorption property and high cationic flocculation, the platelet is adsorbed tothe bottom of the microplate hole; and the indication system comprises IgG anti-D-sensitized O-type red blood cells and a rabbit or goat anti-human globulin reagent. To-be-matched blood plasma is added to the microplate hole already adsorbing a single platelet layer for an immune reaction, and finally through display of the two-step indication system, solid-state method platelet cross-match can bequickly realized. Through creatively using the high-flocculation adsorption effect of polyethyleneimine and using the high-sensitivity two-step indication system, the problems of low sensitivity, high kit price and the like in the current clinical platelet cross-match method can be effectively solved, and a novel kit with a high cost performance is provided for clinical platelet cross-match.

Description

technical field [0001] The invention relates to a kit, which belongs to the field of biotechnology, in particular to a novel platelet cross-matching kit. Background technique [0002] Platelet transfusion, as an effective treatment, has been widely used in clinic. However, due to the long-term blood transfusion, drug treatment and the influence of the disease itself, the patients are prone to produce alloantibodies against platelet antigens in the body. Patient's life safety. In view of this, the National Health and Family Planning Commission stipulated in the "Technical Specifications for Clinical Blood Transfusion" that before platelet transfusion, a cross-matching test must be carried out to improve the quality of blood transfusion for patients and ensure the safety of patients' lives. [0003] The platelet cross-matching method has gone through platelet immunofluorescence test (PIFT), flow cytometry cross-match (FCM), microcolumn gel matching, solid phase agglutination...

Claims

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Application Information

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IPC IPC(8): G01N33/80
CPCG01N33/80
Inventor 罗广平骆宏付涌水姬艳丽叶欣
Owner 广州血液中心