Method and kit for rapidly detecting contents of methylmalonic acid, methyl citric acid, methionine and total homocysteine in dried blood spot

Abstract

The invention discloses a method and a kit for rapidly detecting the content of methylmalonic acid, methyl citric acid, methionine and total homocysteine in dried blood spots. A non-derivatization method is adopted, a UPLC-MS / MS technology is adopted to carry out internal standard method quantitative analysis on a sample, six levels of blood spots are used as correction lines, the concentration ratio of a standard substance solution to an internal standard substance is used as an X axis, and the peak area ratio of the standard substance to the internal standard substance is used as a Y axis; and performing linear regression analysis to obtain a regression equation, substituting the corresponding area of the to-be-detected substance into the regression equation, and calculating the concentration of the to-be-detected substance in the sample. According to the method, methylmalonic acid (MMA), methyl citric acid (MCA), total homocysteine (tHCY) and methionine (MET) can be detected at the same time, the sample size is three blood spots with the diameter of 3.2 mm, sample pretreatment is simple, the specificity is high, time consumed for a single sample is 3 minutes, and high-throughput detection can be achieved.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection, and in particular relates to a rapid detection method and kit for the contents of methylmalonic acid, methylcitric acid, methionine and total homocysteine ​​in dried blood tablets. Background technique [0002] The new screening is based on the abnormal content of methionine and C3 to screen for congenital defects related to the metabolism of propionic acid, methionine, and vitamin B12 (vitamin B12, cobalamin, Cbl). The American Society of Medical Genetics promulgates a new primary screen for differential diagnosis of genetic diseases, mainly 29 congenital genetic diseases, and 25 secondary congenital genetic diseases. Among them, 4 major diseases and 2 minor diseases were related to the abnormal increase of C3 and Met concentrations. [0003] Methylmalonic acidemia (methylmalonic academia, MMA) is an autosomal recessive genetic disease, mainly due to the defect of methylmalonyl-Co...

AI Technical Summary

Problems solved by technology

The derivatization operation can help the liquid phase separation of the isomers methylmalonic acid and succinic acid, and improve the detection sensitivity, but the derivatization pretreatment derivation steps are complicated, the derivatization reagents are toxic and irritating, and the derivatization Due to poor stability and high cost of derivatization reagents, it is necessary to develop a cheap derivatization method with simple pretreatment method, good resolution and stable method

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