Application of AKAP4 protein with 186th-site N-114.04278 in preparing reagent for diagnosing severe oligoasthenospermia

A diagnostic reagent, the technology of less weak sperm, applied in the field of medicine and molecular diagnosis

Active Publication Date: 2017-08-01
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are no reports on non-coding amino acids as biomarkers associated with severe oligoasthenospermia

Method used

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  • Application of AKAP4 protein with 186th-site N-114.04278 in preparing reagent for diagnosing severe oligoasthenospermia
  • Application of AKAP4 protein with 186th-site N-114.04278 in preparing reagent for diagnosing severe oligoasthenospermia
  • Application of AKAP4 protein with 186th-site N-114.04278 in preparing reagent for diagnosing severe oligoasthenospermia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Example 1: Screening of biomarkers associated with severe oligoasthenospermia

[0125] The specific screening method is as follows:

[0126] 1. Sample processing and experimental analysis

[0127]1. Extraction of the whole protein of sperm cells: the same amount of severe oligospermia and normal sperm samples were washed three times with DPBS, and the same amount of RIPA lysate was added to sonicate for 1-2 minutes, incubated on ice for 30 minutes to lyse, and centrifuged at 4°C at 14,000g ×20min to take the supernatant. Protein concentration was determined using the Bradford method.

[0128] 2. Proteolysis: Take approximately 150 μg of sperm protein from the samples of severe oligospermia and normal sperm, use 10% polyacrylamide gel electrophoresis (SDS-PAGE) to separate the proteins, and divide each into 5 parts for enzymatic hydrolysis. Peptides were desalted using ziptip.

[0129] 3. Mass Spectrometry: Nanoflow Liquid Chromatography Separation: Phase A: Water co...

Embodiment 2

[0185] Embodiment 2: clinical detection verification

[0186] Taking 4 healthy samples and 8 clinically diagnosed severe oligoasthenospermia samples as the research objects for verification, the serine at the 208th position of the AKAP3 protein and the mass deviation of -113.05347 at the 186th position of the AKAP4 protein were detected respectively in the above samples. Asparagine shifted, asparagine with mass shift of -114.04278 at position 186 of AKAP4 protein, glutamine with mass shift of -17.02660 at position 617 of AKAP4 protein, lysine with mass shift of +211.09682 at position 733 of AKAP4 protein , lysine with +42.01108 mass shift at position 531 of ATP5A1 protein, lysine with +42.01108 mass shift at position 87 of COX4I1 protein, threonine with +79.96685 mass shift at position 64 of GAPDHS protein and 692 position of KIAA1683 protein The respective detection frequencies of serine with +79.96685 mass shift occurred, and the samples to be tested were diagnosed according...

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Abstract

The invention discloses application of asparagine with AKAP4 protein 186th-site generating mass shift -114.04278 as a biological marker in preparing a reagent for diagnosing severe oligoasthenospermia. The fact that the detection frequency of the AKAP4 protein 186th-site generating mass shift -114.04278 can be used for diagnosing the severe oligoasthenospermia is found, so that a new diagnosis and treatment target is provided for the severe oligoasthenospermia.

Description

technical field [0001] The invention relates to the technical fields of medicine and molecular diagnosis, in particular to the use of asparagine with a mass shift of -114.04278 at position 186 of AKAP4 protein as a marker in the preparation of a diagnostic reagent for severe oligospermia and weak sperm. Background technique [0002] Infertility has become a worldwide reproductive health problem, of which the infertility caused by male factors accounts for about 50%, and it is on the rise in recent years. The main cause of male infertility is oligozoospermia. According to the World Health Organization standard, if the number of a-class sperm is less than 25%, the number of (a+b)-class sperm is less than 50%, and the sperm motility rate is lower than 60%, it can be diagnosed as asthenozoospermia. Oligospermia refers to a condition in which the number of sperm in semen is lower than that of normal fertile men. When a man's sperm is less than 20 million per milliliter, it is ol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N30/88
CPCG01N30/88G01N33/689G01N2030/8818G01N2030/8831G01N2800/367
Inventor 杨静华李翠玲毕白斌陈子江
Owner SHANDONG UNIV
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