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Method for detecting harm degree of postoperative cytomegalovirus infection of kidney transplantation patient

A cytomegalovirus, harmful degree of technology, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve problems that must be treated in time

Pending Publication Date: 2016-02-03
THE FIRST PEOPLES HOSPITAL OF CHANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Specific immune response detection is based on the fact that the complications and organ damage that occur are related to HCMV disease, rather than HCMV replication. Although it is more effective for patients who really need drug treatment, when abnormal immune responses related to HCMV disease are detected , is already in a passive state and must be treated in time

Method used

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  • Method for detecting harm degree of postoperative cytomegalovirus infection of kidney transplantation patient

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Embodiment 1

[0032] This embodiment is a detection method for assessing the degree of harm caused by cytomegalovirus infection for patients with type 1 kidney transplantation after surgery, such as figure 1 shown, including the following steps:

[0033] Step a, blood collection, collecting 20 mL of heparin sodium anticoagulated peripheral blood from a patient undergoing a kidney transplantation.

[0034] Step b, CD19 + Acquisition and analysis of B lymphocytes, mononuclear cells were separated with lymphocyte separation medium (Ficoll) (purchased from Tianjin Haoyang Biological Company), and CD19 in peripheral blood was sorted with magnetic beads + B lymphocytes. Magnetic bead sorting of cells involves the following substeps:

[0035] (1) Take an appropriate amount of mononuclear cell suspension, centrifuge at 100g for 5 minutes, and discard the supernatant;

[0036] (2) by every 10 7 Add 80 μl MACS buffer to each cell, gently blow off the cells, and mix well;

[0037] (3) Add 20 μl ...

Embodiment 2

[0092] This embodiment is a detection method for assessing the degree of harm caused by cytomegalovirus infection in patients with second kidney transplantation after surgery, such as figure 1 shown, including the following steps: .

[0093] Step a, blood collection, collecting 20mL of heparin sodium anticoagulated peripheral blood from patients with second kidney transplantation.

[0094] Step b, CD19 + Acquisition and analysis of B lymphocytes, separation of mononuclear cells with lymphocyte separation medium (Ficoll), separation of CD19 in peripheral blood with magnetic beads + B lymphocytes. Magnetic bead sorting of cells involves the following substeps:

[0095] (1) Take an appropriate amount of mononuclear cell suspension, centrifuge at 100g for 5 minutes, and discard the supernatant;

[0096] (2) by every 10 7 Add 80 μl MACS buffer to each cell, gently blow off the cells, and mix well;

[0097] (3) Add 20 μl CD19 magnetic beads per 80 μl MACS buffer, mix well, and...

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Abstract

The invention relates to a method for detecting harm degree of postoperative cytomegalovirus infection of a kidney transplantation patient. The invention includes the following steps: a, collecting blood; b, acquiring and analyzing CD19+B lymphocytes; c, preparing cytomegalovirus seeds and quantifying; d, culturing the CD19+B lymphocytes in vitro, and using HCMV to directly and indirectly stimulate the CD19+B lymphocytes; e, performing flow detection on expression of a BAFF receptor on the CD19+B lymphocytes; f, detecting apoptosis rate of the CD19+B lymphocytes; g, detecting IgG secretion capability of the CD19+B lymphocytes. By the method, reaction, to HCMV stimulation, of the B lymphocytes of the patient can be analyzed. The method has important value in evaluating chronic rejection which is indirect effect caused by HCMV infection of the kidney transplantation patient.

Description

technical field [0001] The invention relates to a detection method for assessing different harm levels caused by cytomegalovirus infection in kidney transplant patients after operation. Background technique [0002] Human cytomegalovirus (HCMV) is a DNA double-stranded virus that widely exists in the human population and is an opportunistic pathogen. HCMV infection is the most common infection in transplant patients or other immunocompromised patients treated with immunosuppressants. Initial infection with HCMV in immunocompetent individuals is usually asymptomatic, but the severity of infection in transplant patients is proportional to the degree of immunosuppression, and HCMV infection is the most common viral infection complication in kidney transplant patients. [0003] Transplanted solid organs are the main organs of HCMV infection. The clinical symptoms caused by HCMV infection in transplant patients can be divided into two categories: direct effects (monocytosis synd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N5/0784
Inventor 徐海燕何小舟马旭怡宋丹
Owner THE FIRST PEOPLES HOSPITAL OF CHANGZHOU
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