Marker for detecting and evaluating liver regeneration and application thereof

Abstract

The invention also discloses application of CD63, albumin ALB and endothelial growth factor VEGF or specific antibody thereof as markers in preparation of a kit for detecting and evaluating liver regeneration. The marker CD63 + ALB + VEGF for noninvasively detecting and evaluating the liver regeneration condition by using peripheral blood is provided for the first time, the sensitivity of the marker provided by the invention in liver failure liver regeneration evaluation is as high as 87.5%, the specificity is as high as 90%, and the marker is closely associated with liver regeneration. In a liver failure patient, the quantity of exosomes marked with CD63 + ALB + VEGF is significantly higher in the survival group than in a death group. Results show that the exosome with VEGF and ALB in blood plasma is a more accurate and specific marker for liver regeneration, and the detection technology may become a new non-invasive diagnosis technology for liver regeneration.

Description

technical field [0001] The invention relates to a biochemical marker, in particular to a marker and a detection kit for detecting and evaluating liver regeneration, and belongs to the technical field of medical testing. Background technique [0002] There are about 350 million chronically infected people with hepatitis B virus (HBV) in the world, and my country accounts for one-third of them (about 120 million). Chronic HBV infection gradually develops into severe hepatitis and the liver failure caused by it is the most dangerous and the worst clinical type of hepatitis, with a fatality rate of about 40% to 60%. Because liver failure often affects young adults, with an average age of onset of about 45 years, it is very harmful to individuals, families and society. One of the main differences between HBV-related liver failure and end-stage cirrhosis is the regenerative potential of the liver. Patients with end-stage cirrhosis who do not undergo liver transplantation end up w...

AI Technical Summary

Problems solved by technology

[0005] However, there are many problems in traditional exosome extraction methods, which limit the application of exosomes in clinical testing.

For example, the exosomes extracted by the kit extraction method contain other vesicles and macromolecular impurity proteins, which affect the purity of exosomes.

Gel exclusion chromatography requires specialized equipment, is time-consuming and not widely used

Immunomagnetic bead method is inefficient, the antibody is expensive and cannot be recovered, and it is difficult to be widely used

Although the traditional centrifugation method is cumbersome, it is still the most commonly used method for exosome extraction.

The existing differential centrifugation method has relatively simple requirements for experimental equipment, which can basically meet the extraction requirements, but its existing problems are: (1) The yield of exosomes is low

Flow cytometry cannot detect the target results, because traditional flow cytometers are difficult to detect particles with a diameter below 1um, and only Amnis can be used to quantify and analyze flow cytometers. The sensitivity and image resolution of Amnis can detect diameters The exosomes range from 30 to 100nm, but the minimum detection parameters of Amnis quantitative imaging analysis flow cytometer are that the sample particle flow rate reaches at least 100 obj / s, the total number of collected particles is at least 20,000, and the exosome yield obtained by existing extraction methods Too low, it is difficult to meet the detection requirements

(2) The extraction steps and methods have a certain impact on the physical and chemical properties of exosomes. For example, the integrity of exosomes cannot be guaranteed. Existing techniques can easily cause exosomes to rupture and affect their functionality.

[0006] In addition, although there are many exosome markers, experiments have proved that many markers are not suitable for detection of liver regeneration, and the labeling effect is not good, which directly affects the test results

Images