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Kit for detection of oligospermia

A detection reagent and technology for oligospermia, applied in the field of kits for detection of oligospermia, can solve the problems of incomplete maturation of spermatogenic epithelium, affecting sperm motility, hindered testicular development, etc. High, low false positive effect

Active Publication Date: 2019-02-01
广州市普森生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

③ Lack of trace elements affects sperm motility
④ The development of the testis is blocked, and the seminiferous epithelium of the testis is not fully mature or damaged, resulting in poor sperm quality and weakened vitality
⑤Insufficient semen volume affects sperm motility

Method used

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  • Kit for detection of oligospermia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Detection of CRKL gene mutation and its association with oligospermia

[0035] 1.1 Research object

[0036] A total of 100 patients with oligospermia were selected and diagnosed and treated in the First Affiliated Hospital of Soochow University from July 2012 to December 2017. The clinical data were obtained from medical records. 50 age-similar controls were healthy people who underwent physical examination in the physical examination center. Take 1.5ml of peripheral blood from the test personnel and store it in a freezer at -80°C. All subjects obtained consent in accordance with the requirements of the Ethics Committee of the First Affiliated Hospital of Soochow University.

[0037] 1.2 DNA extraction

[0038] The peripheral blood DNA of the subjects was extracted by the conventional phenol-chloroform method, as follows:

[0039] 1) Take 1ml of peripheral blood, centrifuge at 1600g at 3-6°C for 8-12min, centrifuge the supernatant at 16000g at 4°C for 8-12...

Embodiment 2

[0052] Embodiment 2 Typing result verification

[0053] Since the mutation of CRKL81 is highly related to oligospermia, gene-specific primers can be designed based on this mutation and then expanded detection using the patient's DNA as a template.

[0054] Prepare a kit (100 person-times), the composition of the kit is: upstream and downstream primers, dNTP, Mg2+, Taq enzyme, PCR buffer, the reaction system is 20 microliters, upstream primer: 5'-ctctggaaattcgttaatgt-3'; downstream primer: 5 '-gtccacaagggagagcagac-3'.

[0055] Taking the sample in Example 1 as the detection object, re-perform PCR and result sequencing, and select positively associated SNPs by comparing the genotype distribution frequency, and use the regression coefficient of a single SNP in the whole genome scan sample as the weight to further obtain the risk score Value, draw ROC to evaluate the sensitivity and specificity of prediction, and then evaluate the ability of these SNPs to judge the onset of oligo...

Embodiment 3

[0057] Embodiment 3 oligospermia detection accuracy verification

[0058] 2ml of peripheral blood was drawn from the subjects (20 newly identified oligospermia patients and 10 healthy controls) were extracted, and DNA was extracted by conventional methods. Use the oligospermia susceptibility detection kit for PCR reaction, PCR amplification conditions: 94°C for 5 minutes, (94°C for 35 seconds, 65°C for 30 seconds, 72°C for 45 seconds) × 30, 72°C for 10 minutes, 10°C for incubation . The reaction products were sequenced, and the sequencing results were checked and analyzed using Meglign 7.0 and Chromas 2.33 software. 95% of the subjects with CRKL81t>c mutation in the detection results were oligospermia patients, and the mutation was not detected in 10 control cases, and the accuracy was high.

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Abstract

The invention discloses a kit for detection of oligospermia. The kit contains detection reagents capable of detecting an oligospermia related molecular marker, and specifically the related detection site is a CRKL gene fragment, when t / c mutation exists in the sequence, an oligospermia patient can be diagnosed. The kit provided by the invention has the advantages and beneficial effects that designed specific primers are utilized and the PCR technology is adopted, therefore the kit has very high detection sensitivity, high accuracy, good specificity and low false positive, is simple in operation, safe, high in automation degree, and anti-pollution, also has fast speed and high flux, can be completed in 2-3h, is economical and convenient, and is easy for wide promotion in clinical practice,saves patients' expenditure, and can achieve batch detection.

Description

field of invention [0001] The invention relates to the technical field of detection, in particular to a kit for detecting oligospermia. Background technique [0002] Oligozoospermia refers to poor or even inactive sperm motility. In men, oligozoospermia is one of the main causes of male infertility or decreased fertility. There are many reasons for asthenozoospermia. Whether it causes infertility depends on the degree of asthenospermia. If it is mild asthenozoospermia, it generally does not have a big impact on fertility. If it is severe asthenozoospermia, further examinations are needed to find out the cause of the asthenospermia. Is there any asthenospermia caused by prostatitis? Asthenozoospermia caused by immune factors, asthenospermia caused by asthenozoospermia is found out and treated according to the cause. [0003] Oligozoospermia and asthenozoospermia are mainly caused by the following causes: ① Inflammation of epididymis, seminal vesicles, prostate and other or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 王振张勇
Owner 广州市普森生物科技有限公司
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