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Chromatographic column, and kit and device for detecting prosthetic joint infection

A chromatographic analysis and chromatographic column technology, applied in the direction of measuring devices, analytical materials, material separation, etc., can solve problems such as inability to achieve instant diagnosis, no effective solution, and insufficient sensitivity, so as to improve stability and separation Good, the effect of improving sensitivity

Active Publication Date: 2019-08-20
北京谷海天目生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These indicators still have some deficiencies in the specificity and sensitivity of diagnosing 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) Counting 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 Care Testing, POCT, or bedside diagnosis)
[0004] Therefore, how to detect PJI more accurately, there is still no effective solution in the prior art

Method used

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  • Chromatographic column, and kit and device for detecting prosthetic joint infection
  • Chromatographic column, and kit and device for detecting prosthetic joint infection
  • Chromatographic column, and kit and device for detecting prosthetic joint infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] The heavy isotope-labeled standard peptides shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 were prepared with a final concentration of 100fmol of peptides, then shake and mix, centrifuge for 3 minutes on a 12000g centrifuge, and take the supernatant for later use;

[0098] Extract protein from 40 joint fluid samples from individuals with prosthetic joints, and then perform enzymatic hydrolysis. Take 100 μg of joint fluid proteolytic peptides from each sample and add 100 μL of sample buffer, shake and mix to prepare Joint fluid peptide solution for standby;

[0099] Take 1 μL of joint fluid peptide solution, and add 1 μL of 100 fmol heavy isotope-labeled standard peptide solution to it, then add 3uL of loading buffer, mix well, and place on a 96-well loading plate for chromatographic injection.

[0100] Mix the synthesized heavy-labeled peptides in an equimolar ratio, and mix the individual peptides according to the following concentrations: 250fmol,...

Embodiment 2

[0123] The difference between Example 2 and Example 1 is that in the chromatographic analysis column, the length of the chromatographic column tube is 150 mm, the outer diameter is 360 μm, and the inner diameter is 150 μm, and the filler is C18 reverse phase filler with a particle size of 1.9 μm. Meanwhile, the chromatographic separation time was 75 minutes.

Embodiment 3

[0125] The difference between Example 3 and Example 1 is that in the chromatographic analysis column, the length of the chromatographic column tube is 300 mm, the outer diameter is 360 μm, and the inner diameter is 150 μm, and the filler is C18 reverse phase filler with a particle size of 1.9 μm. Meanwhile, the chromatographic separation time was 150 minutes.

[0126] Both Examples 2 and 3 can effectively identify the target peptide, and carry out targeted quantitative analysis of the target peptide, and the results obtained can also well distinguish whether the prosthetic joint is infected or not (due to the data in Example 1 There is no significant difference and will not be specifically listed here). However, considering the timeliness of clinical sample detection and the cost of mass spectrometry detection, the detection conditions in Example 1 in this application are the best application conditions.

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Abstract

The present invention provides a chromatographic column, and a kit and a device for detecting prosthetic joint infection. The chromatographic column comprises: a chromatographic column tube, packing filled in the chromatographic column tube, and a chromatographic column tip integrated with the chromatographic column tube, wherein the inner diameter of the chromatographic column tube is 100-200 [mu]m. By using a relatively wide inner diameter, the column pressure of the chromatographic column tube can be maintained within the normal pressure range while increasing the chromatographic elution flow rate, so that the stability of chromatographic separation is improved, the sample loading volume of the sample can be increased in a facilitated manner, and sensitivity of mass spectrometry can beimproved. By using the integrated chromatographic column tip, there is no tailing phenomenon when performing elution, and the separation degree of the protein is better. Therefore, according to the technical scheme of the present invention, when the PIJ is detected by using the improved chromatographic column, the peptide has a better separation degree and higher stability, and the quantitative result of the target protein can be more accurate.

Description

technical field [0001] The invention relates to the field of prosthetic joint infection detection, in particular to a chromatographic analysis column, a kit and a device for detecting prosthetic joint infection. Background technique [0002] Prosthetic joint infections (PJI) is recognized as one of the most serious complications after joint replacement. Although the incidence of PJI is not high, it has not been significantly reduced since the application of antibiotics. The reported incidence of hip and shoulder PJI is approximately 1%, knee PJI incidence is approximately 2%, and elbow PJI incidence is approximately 9%. For arthroplasty, the incidence of PJI is even as high as 40%; in addition, 33% of the complications leading to the failure of total ankle arthroplasty are PJI. [0003] 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 c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/38
CPCG01N30/88G01N30/38G01N2030/8831
Inventor 秦钧陈继营汪宜丁琛宋雷李睿李恺刘明伟
Owner 北京谷海天目生物医学科技有限公司
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