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CN-122017219-A - Alzheimer's disease jugular deep vein lymphatic anastomosis curative effect prediction and adaptive population screening kit and application

CN122017219ACN 122017219 ACN122017219 ACN 122017219ACN-122017219-A

Abstract

The application relates to the field of in-vitro diagnosis kits, and discloses a kit for predicting curative effect of deep jugular vein lymphatic anastomosis of Alzheimer disease and screening adaptive population and application thereof. The kit comprises a mass spectrum detection module, an ELISA detection module and a logistic regression model, wherein the mass spectrum detection module is used for detecting the NPTX2 and IGSF10 protein concentration in a sample, the ELISA detection module is used for detecting ptau and Abeta 42 protein concentration in the sample, and the LVA operation curative effect is predicted and the adaptive population is screened by calculating the NPTX2/IGSF10 ratio and the ptau/Abeta 42 ratio and combining the logistic regression model. The kit provided by the application has high sensitivity and specificity, can accurately detect the concentration of related proteins in a sample, and provides a reliable basis for the curative effect prediction of Alzheimer's disease deep jugular vein lymphatic anastomosis (LVA) and the screening of adaptation groups.

Inventors

  • YU ZHAOYAN
  • XIAO JINRONG

Assignees

  • 山东大学

Dates

Publication Date
20260512
Application Date
20260128

Claims (10)

  1. 1. The kit for predicting the curative effect of the deep jugular vein lymphadenitis operation of the Alzheimer's disease and screening the adaptive population is characterized by comprising a mass spectrum detection module, an ELISA detection module and a logistic regression model; The mass spectrum detection module is used for detecting the NPTX2 and IGSF10 protein concentration in a sample; the ELISA detection module is used for detecting ptau and Abeta 42 protein concentrations in a sample; And (3) predicting the LVA operation curative effect and screening the adaptive population by calculating the NPTX2/IGSF10 ratio and ptau/Abeta 42 ratio and combining a logistic regression model.
  2. 2. The kit of claim 1, wherein the test sample of the kit is a cerebrospinal fluid sample.
  3. 3. The kit of claim 2, wherein the logistic regression model is: logit(P) = 1.807×(NPTX2/IGSF10) + 10.461×(ptau181/Abeta42) – 3.172; Wherein, P is the probability of clearance of toxic protein in cerebrospinal fluid after operation, and when P is more than or equal to 0.355, the patient is judged to be LVA operation adaptation group.
  4. 4. The kit of claim 1, wherein the mass spectrometry detection module comprises: mobile phase A is 0.1% formic acid aqueous solution, and B is 0.1% formic acid acetonitrile solution; A redissolution reagent, which is 0.1% formic acid solution and contains an internal standard 13 C marked NPTX2 and IGSF10 peptide fragment; standard substances are NPTX2 and IGSF10 standard substances, and the concentration gradients are 0.05, 0.1, 1, 5 and 10 ng/mL; consumable 0.22 μm organic phase filter membrane and C18 reversed phase chromatographic column.
  5. 5. The kit of claim 4, wherein the mass spectrometry detection module screens the characteristic peptide fragments using DIA mode and targets the characteristic peptide fragments by MRM mode, and the ion pairs are NPTX2 m/z 794.19 →312.1 and IGSF10 m/z 797.58 → 295.0.
  6. 6. The kit of claim 1, wherein the ELISA detection module comprises: pre-coating antibody plates, namely coating anti-ptau and anti-Abeta 42 monoclonal antibodies; The detection reagent comprises enzyme-labeled secondary antibody, substrate solution and stop solution; Standard substances ptau and Abeta 42, the concentration gradients are 10, 20, 40, 80 and 160 pg/mL; Auxiliary reagents, namely a washing liquid and a sealing liquid.
  7. 7. The kit according to claim 1, wherein, the kit further comprises a cerebrospinal fluid pretreatment group comprising: Pretreatment reagents for mass spectrometry, namely DDM lysate, DTT solution, IAA solution, trypsin and pH8.0 Tris-HCl buffer solution; pretreatment reagent for ELISA, diluent.
  8. 8. The kit of claim 7, wherein the cerebrospinal fluid sample dilution ratio in the ELISA assay is 1:10.
  9. 9. The kit according to claim 1, wherein, the kit further comprises a quality control and auxiliary group, wherein the quality control and auxiliary group comprises: quality control material, which is cerebrospinal fluid matrix quality control liquid containing NPTX2, IGSF10, ptau181 and Abeta 42 with known concentration and blank cerebrospinal fluid; auxiliary data, cerebrospinal fluid collection guidelines, detection specifications, and logistic regression model calculation tables.
  10. 10. Use of a kit according to any one of claims 1-9 for the preparation of a predictive therapeutic effect of deep jugular lymphadenitis in alzheimer's disease and for the screening of products for adaptation populations.

Description

Alzheimer's disease jugular deep vein lymphatic anastomosis curative effect prediction and adaptive population screening kit and application Technical Field The application relates to the field of in-vitro diagnostic kits, in particular to a kit for predicting curative effect of deep jugular lymphatic anastomosis of Alzheimer disease and screening adaptive population and application thereof. Background Alzheimer's Disease (AD) is a globally highly neurodegenerative disease with progressive cognitive decline, memory decline as a central symptom, pathologically characterized by abnormal deposition of amyloid (Abeta) in the brain to form senile plaques, aggregation of hyperphosphorylated tau protein (ptau) to form neurofibrillary tangles, and synaptic injury and neuronal loss. The cerebrospinal fluid is used as a gold sample for AD detection, has the irreplaceable advantages of directly reflecting the pathological state in brain, namely, the cerebrospinal fluid is directly communicated with interstitial fluid of brain tissue, wherein the concentrations of markers such as Abeta 42, ptau181, NPTX2 and the like are highly synchronous with the pathological progress in brain, and the marker has more specificity than peripheral blood (such as plasma), the concentration of AD related protein in the cerebrospinal fluid is obviously higher than that in the peripheral blood (such as ptau181 in the cerebrospinal fluid is 20-80pg/mL, and the peripheral blood is only 1-5 pg/mL), and can be detected without complex enrichment. Deep jugular lymphatic anastomosis (LVA) is a microsurgery procedure, and is used for assisting treatment of AD at present by anastomosing a superficial cervical lymphatic vessel with a deep jugular vein, reconstructing a lymphatic drainage channel of a brain and promoting clearance of neurotoxic proteins such as aβ, ptau and the like in the brain. However, the clinical application of LVA surgery has the bottleneck of difficult screening of the key adaptive population, and the lack of objective indexes can judge which AD patients can benefit from the surgery, and the postoperative efficacy of partial patients is poor. The existing AD molecular detection technology cannot meet clinical requirements of LVA operation, and the specific problems comprise that the existing AD diagnosis depends on a cognition evaluation scale (such as a simple mental state check list MMSE), imaging examination (such as skull MRI/PET) and a single molecular marker (such as cerebrospinal fluid ptau and Abeta 42), the cognition scale is high in observability (1), the cognition scale is greatly influenced by patient coordination degree and evaluator experience, (2) the accuracy is insufficient, the prediction capability of the single marker (such as ptau 181/Abeta 42) on AD progress and LVA operation treatment response is limited, (3) the technical cooperativity is poor, or only a small amount of protein (which is covered narrowly, is difficult to detect NPTX2/IGSF 10) by mass spectrum detection, or only an ELISA detection ptau/Abeta 42 (which lacks operation target related protein data) is not formed, and a cerebrospinal fluid special complementation system of 'mass spectrum+ELISA' is lacked, and the ratio of the multiple markers (such as NPTX2/IGSF10 and ptau/Abeta 42) is not combined with clinical curative effect to construct a mathematical model, and the probability of the improvement and the operation after the improvement cannot be quantified. In summary, the existing AD molecular detection technology cannot meet the clinical requirements of LVA surgery. Disclosure of Invention The invention aims to provide a high-precision modularized joint detection kit special for cerebrospinal fluid, which detects NPTX2/IGSF10 in cerebrospinal fluid by high-resolution mass spectrometry, detects ptau/Abeta 42 in cerebrospinal fluid by ELISA, and combines a logistic regression model to realize accurate pre-operation screening of LVA operation adaptation groups. In order to achieve the above object, the present application adopts the following technical scheme: In a first aspect, the application provides a kit for predicting curative effect of deep jugular vein lymphadenitis of Alzheimer disease and screening adaptive population, wherein the kit comprises a mass spectrum detection module, an ELISA detection module and a logistic regression model; The mass spectrum detection module is used for detecting the NPTX2 and IGSF10 protein concentration in a sample; the ELISA detection module is used for detecting ptau and Abeta 42 protein concentrations in a sample; And (3) predicting the LVA operation curative effect and screening the adaptive population by calculating the NPTX2/IGSF10 ratio and ptau/Abeta 42 ratio and combining a logistic regression model. Further, the detection sample of the kit is a cerebrospinal fluid sample. Further, the logistic regression model is: logit(P) = 1.807×(NPTX2/IGSF10) + 10.461×(ptau181/Abeta42) – 3.172; Wherei