Method and kit for detecting metabolites in dried blood spots

A metabolite and dried blood film technology, applied in the field of biochemical detection, can solve problems such as easy to produce false positive results, achieve high diagnostic efficiency, reduce diagnosis time, and reduce false positive rates

Active Publication Date: 2019-01-04
GENERAL HOSPITAL OF PLA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the screening of these four diseases is mainly based on liquid chromatography-tandem mass spectrometry. The existing detection kits do not have these specific indicators, and can only show methionine and propionyl meat, which are dependent on methionine MET and propionate metabolism. Increased base C3, however, these analytes are not specific for the diagnosis of MMA and PA and are prone to false positive results

Method used

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  • Method and kit for detecting metabolites in dried blood spots
  • Method and kit for detecting metabolites in dried blood spots
  • Method and kit for detecting metabolites in dried blood spots

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Embodiment Construction

[0037] The kit and detection method for detecting metabolites in dried blood slices of the present invention will be described in detail below in conjunction with the examples.

[0038] Instruments used:

[0039] Ultra-high performance liquid chromatography-tandem mass spectrometer (Waters ACQUITY UPLC I-Class IVD / Waters XevoTQ-D IVD System, USA);

[0040]Nitrogen blowing ns instrument (model AD 8001, Mass Spec Biotechnology Co., Ltd.);

[0041] Microplate shaker (model OST7701, Mass Spec Biotechnology Co., Ltd.);

[0042] V cut-off 96-well plate (Product No. Z011075, Mass Spec Biotechnology Co., Ltd.);

[0043] V-bottom 96-well plate (Product No. Z011076, Mass Spec Biotechnology Co., Ltd.);

[0044] 1 / 8 inch (3.2mm) hole punch;

[0045] Kit components:

[0046] (1) Extractant: methanol water (80:20), containing 0.02–0.05% formic acid.

[0047] (2) Derivatization reagent: 3N n-butanol hydrochloride;

[0048] (3) Complex solution: ultrapure water, containing 0.02–0.05% f...

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Abstract

The invention relates to a method and a kit for detecting metabolites in dried blood spots. The metabolites include malonic acid, methyl malonic acid, ethyl malonic acid, me-citrate and total homocysteine, extraction agents containing internal standards are added into the dried blood spots to perform extraction, vibration incubation and derivatization on the dried blood spots, and the malonic acid, the methyl malonic acid, the ethyl malonic acid, the me-citrate and the total homocysteine in treated dried blood spot samples are detected by a liquid chromatography-tandem mass spectrometry method. The concentration of the malonic acid, the methyl malonic acid, the ethyl malonic acid, the me-citrate and the total homocysteine in the filter paper dried blood spot samples can be measured, the method and the kit are used for auxiliary diagnosis of methylmalonic acidemia, methylmalonic acidemia combined homocystinemia, homocystinemia and propionic acidemia, diagnosis efficiency is high, and diagnosis time can be shortened.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection, in particular to a method for detecting the concentration of metabolites such as malonic acid, methylmalonic acid, ethylmalonic acid, methylcitric acid and total homocysteine ​​in dried blood tablets Methods and kits. Background technique [0002] Methylmalonic acidemia is an autosomal recessive genetic disease, first reported in 1967 by Oberholzer et al. The disease is mainly due to the defect of methylmalonyl-CoA mutase itself or its coenzyme cobalamin metabolism defect, resulting in the metabolism of methylmalonic acid, 3-hydroxypropionic acid, malonic acid and methyl citrate. Abnormal accumulation of substances, causing relevant clinical manifestations. The age of onset of methylmalonic acidemia varies. Early-onset children with onset within 1 year of age have a poor prognosis and are easily missed or misdiagnosed. The mortality and disability rates are high. Delayed treatment...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 张春燕田亚平
Owner GENERAL HOSPITAL OF PLA
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