Application of synovial fluid neutrophil gelatinase-associated lipocalin in detection and diagnostic kit for prosthetic joint infection

A lipocalin, neutrophil technology, applied in the field of medical diagnosis, can solve the problems of high price, low specificity, low sensitivity, etc., and achieves good application and industrialization prospects, strong detection sensitivity, good The effect of anti-interference ability

Inactive Publication Date: 2018-09-28
THE FIRST AFFILIATED HOSPITAL OF FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are still some deficiencies in the specificity and sensitivity of these indicators in the diagnosis of PJI: 1. Bacterial culture is one of the "gold" standards for diagnosing PJI, but the positive rate of bacterial culture is about 60% to 70%, and the sensitivity is not high enough
2. The count and classification of white blood cells in synovial fluid is an effective method for diagnosing PJI, but after joint replacement in patients with systemic inflammatory arthritis, this index will also increase, and the specificity is not strong
3. Existing laboratory diagnostic methods cannot achieve immediate diagnosis (Point of CareTesting, POCT, or bedside diagnosis)
Enzyme-linked immunosorbent assay (ELISA) kits are mostly used for the detection of existing synovial fluid biomarkers, and a certain number of specimens need to be collected clinically for detection, which is expensive and cannot meet the needs of clinical individualization and POCT
Colloidal gold immunolabeling technique is a semi-quantitative technique, which is easy to use but cannot be quantitatively detected

Method used

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  • Application of synovial fluid neutrophil gelatinase-associated lipocalin in detection and diagnostic kit for prosthetic joint infection
  • Application of synovial fluid neutrophil gelatinase-associated lipocalin in detection and diagnostic kit for prosthetic joint infection
  • Application of synovial fluid neutrophil gelatinase-associated lipocalin in detection and diagnostic kit for prosthetic joint infection

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 Preparation of joint fluid NGAL immunofluorescence test strip

[0023] 1. Antibody preparation

[0024] 1) Prepare anti-NGAL antibody and goat anti-mouse IgG antibody by genetic engineering method; the antibody can be prepared by the laboratory itself, or can be entrusted to a commercial company.

[0025] 2) Select time-resolved fluorescent latex particles, and screen them in terms of particle size and surface groups; for example, polymer polystyrene particles with a diameter of 100nm, embedded with europium-chromium compounds, have the characteristics of time-resolved fluorescence, and the surface of the particles It has a high-density functional group-COOH (the above-mentioned particles and their parameters are not the limiting conditions of the present invention); it can be purchased directly from commercial companies.

[0026] 3) Fluorescent particle-labeled antibody, using the covalent coating method: take 25 mg of particles and wash them twice with P...

Embodiment 2

[0035] The collection of embodiment 2 joint fluid specimen and clinical data

[0036] Approved by the Ethics Committee of the First Affiliated Hospital of Fujian Medical University, the synovial fluid of patients re-admitted for various reasons after artificial joint replacement was collected. At the same time, detailed clinical data of patients were collected. According to the diagnostic criteria of the American Musculoskeletal Infection Society (MSIS), they were divided into an infection group of 22 cases and a non-infection group of 23 cases. After centrifuging the joint fluid, the supernatant was collected and frozen for future use. Before use, thaw the stored joint fluid and stabilize at room temperature for later use.

Embodiment 3

[0037] Embodiment 3 adopts ELISA method to detect synovial fluid NGAL

[0038] 1) Take 1 mL of joint fluid and centrifuge at 8000rpm for 4 minutes to separate the supernatant and cells of the joint fluid, absorb the supernatant, transfer it to an EP tube, and store it in a -80°C refrigerator for testing.

[0039] 2) According to the ELISA kit instruction method, use the ELISA kit (CUSABIO company) to detect NGAL in the supernatant, detect the content of NGAL in the joint fluid of patients in the infection group and non-infection group, and perform statistical analysis.

[0040] 3) The clinical data of 45 patients were collected, and the threshold value for diagnosing PJI was determined according to the diagnostic criteria of MSIS.

[0041] 4) The experimental results are as follows: the median of NGAL expression in the infection group was 221.9ng / mL (P25=138.1ng / mL, P75=305.7ng / mL), and the median of NGAL expression in the non-infection group was 39.5ng / mL mL (P25=12.2ng / mL, ...

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Abstract

The invention relates to an application of synovial fluid neutrophil gelatinase-associated lipocalin (NGAL) in detection and a diagnostic kit for prosthetic joint infection. Joint replacement is an effective treatment mode for treating joint disease at a terminal stage, and periprosthetic joint infection is a severe complication after joint replacement. The synovial fluid NGAL has high sensitivityand specificity for diagnosing the prosthetic joint infection. According to the invention, fluorescence particles are taken as a maker, and the diagnostic kit (a test paper strip) for the synovial fluid NGAL is prepared through an immunochromatography method. By using an immunofluorescence quantitative analyzer, quantitative detection and instant detection on a synovial fluid specimen can be carried out, and diagnosis of the prosthetic joint infection is realized. A synovial fluid NGAL detection reagent and the diagnostic kit have the advantages of simple usage, sensitive reaction, and high diagnosis accuracy.

Description

technical field [0001] The invention relates to the field of medical diagnosis, in particular to the application of synovial fluid neutrophil gelatinase-associated lipocalin (NGAL) in detection of artificial joint infection and a diagnostic kit. Background technique [0002] Artificial joint replacement is an effective method to treat severe joint diseases, but artificial joint infection (Prosthetic Joint Infection, PJI) is one of the serious complications after joint replacement, and its incidence is about 1% to 2% in the initial joint replacement surgery , Revision surgery up to 3% to 10%. The clinical manifestations of PJI are complex, and there are still many difficulties in its diagnosis and treatment. According to the diagnostic guidelines proposed by the American Academy of Orthopedic Surgeons, the Infectious Diseases Society, and the Musculoskeletal Infection Society, most PJIs can be clearly diagnosed and treated promptly. However, there are also some patients wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558
CPCG01N33/558G01N33/6803G01N2333/47
Inventor 张文明李文波康加祥洪海峰白国昌汪大明张翀景黄子达李孟庆方心俞张超凡肖江群胡德庆
Owner THE FIRST AFFILIATED HOSPITAL OF FUJIAN MEDICAL UNIV
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