Method and kit for detecting various related substance of CAH at same time

A technology of related substances and kits, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of lack of efficient detection kits, improve screening efficiency and screening quality, improve screening quality, and improve screening efficiency effect

Inactive Publication Date: 2018-05-29
SHENZHEN HUADA GENE INST
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  • Method and kit for detecting various related substance of CAH at same time
  • Method and kit for detecting various related substance of CAH at same time
  • Method and kit for detecting various related substance of CAH at same time

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Embodiment

[0047] The kit for this example includes the following components individually packaged:

[0048] (A) Mixed isotope internal standard, 1 bottle, the weight of 1 bottle of mixed isotope internal standard is 100ng; including 25ng of 17α hydroxyprogesterone isotope internal standard 13 C 3 -17αOHP, 25ng of androstenedione isotope internal standard, 12.5ng of 11-deoxycortisol isotope internal standard, 12.5ng of 21-deoxycortisol isotope internal standard and 25ng of cortisol isotope internal standard;

[0049] (B) Standard curve product, 1 set; the standard curve product is human whole body of five hormones containing 17α hydroxyprogesterone, androstenedione, 11-deoxycortisol, 21-deoxycortisol and cortisol with known gradient concentration Dried blood film of blood filter paper, a set of 8 sheets in total, marked as cal1 to cal8;

[0050] Cal1 contains 0.5 ng / mL of 17α-hydroxyprogesterone, 0.5 ng / mL of androstenedione, 0.5 ng / mL of 11-deoxycortisol, 0.5 ng / mL of 21-deoxycortisol...

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Abstract

The invention discloses a method and a kit for detecting various related substance of CAH at the same time. The method is used for detecting 17 alpha progesterone, androstenedione, 11-deoxycortisol, 21-deoxycortisol and cortisol at the same time. The method includes: (1), adding an organic solvent into a mixed internal standard product to prepare a stock solution, and diluting to obtain extractionworking liquid; (2), using the extraction working liquid to incubate a to-be-detected blood spot sample; (3), centrifuging, taking supernate, concentrating, and using 45% methanol for re-dissolving;(4), using LC-MS/MS to detect a re-dissolving product, wherein the mixed internal standard product includes isotope internal standard products of 17 alpha progesterone, androstenedione, 11-deoxycortisol, 21-deoxycortisol and cortisol. The method can quantify five hormones in one time, can provide specific, sensitive, low-false positive and high-positive-prediction-value CAH screening and can screen CAH of 11-OHD and 21-OHD at the same time, thereby improving screening efficiency and quality.

Description

technical field [0001] The present application relates to the field of detection of congenital adrenal hyperplasia-related substances, in particular to a method and a kit for simultaneously detecting multiple CAH-related substances. Background technique [0002] Congenital adrenal hyperplasia (abbreviated as CAH) is a group of autosomal recessive genetic diseases caused by the lack of certain enzyme activity in the process of adrenal corticosteroid hormone synthesis and metabolism, which leads to cortisol synthesis disorder. The main clinical manifestation is adrenal cortex hypofunction, Some children are accompanied by electrolyte imbalance and abnormal gonadal development, which may be life-threatening due to adrenal crisis in the neonatal period. CAH is divided into 6 types according to enzyme deficiency, as shown in Table 1, the most common of which is 21-hydroxylase deficiency (21-OHD), accounting for 90% to 95%, followed by 11-hydroxylase deficiency ( 11-OHD), account...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/02
CPCG01N30/02G01N30/06
Inventor 廖云莉饶维桥叶文利陈晓敏卢鹏飞林梁訾金
Owner SHENZHEN HUADA GENE INST
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