Acrosome reaction kit, method for inducing acrosome reaction and detection and method for evaluating acrosome reaction condition

Abstract

The invention discloses an acrosome reaction kit, which comprises tissue slide preserving fluid, a fluorescent conjugate and a fluorescent sealing solid liquid, wherein each 100 ml of the tissue slide preserving fluid contains 0.5ml of glutaraldehyde, 5ml of methanol, 2ml of ethanol, 0.2g of polyvinylpyrrolidone (PVP), 1ml of formalin and the balance of purified water. The kit can carry out a sperm-induced acrosome reaction and detection; and if calcium ion induction is not carried out, the kit can carry out the acrosome reaction condition evaluation and the acrosome integrity determination. By the tissue slide preserving fluid, the test process for the sperm-induced acrosome reaction and detection is carried out by two parts, samples and test results which are obtained in the first part can be preserved and the final test result cannot be influenced if the operations of the second part are carried out in the subsequent 72 hours, so that the work intensity can be relieved. The fluorescence of a fluorescent staining system is strong and cannot be attenuated after being stabilized for 15 days, so that the detection result is more stable and is easy to interprete and the repeatability of the result can be ensured. The kit and a determination method can be clinically applied and popularized.

Description

technical field [0001] The present invention relates to sperm acrosome function screening, especially simulating the fertilization environment in the human body in vitro, initiating sperm capacitation and inducing acrosome reaction in vitro, and evaluating the ability of capacitated sperm to undergo acrosome reaction by fluorescent staining, thereby evaluating The ability of sperm to penetrate oocytes and fertilize them provides a theoretical basis for the analysis of the etiology of infertility and the selection of assisted reproductive treatment options. technical background [0002] Male sex and its associated factors account for at least 50% of the causes of infertile couples. In clinical practice, male fertility is mainly evaluated by routine semen analysis that detects indicators such as sperm density, motility, and morphology. However, the indicators provided by routine semen analysis only reflect the quality of sperm, and only the function of sperm can reflect the in...

AI Technical Summary

Problems solved by technology

[0004] At present, sperm acrosome function tests include sperm morphology and integrity test, zona pellucida-induced sperm acrosome reaction test, etc. Intra-injection) has a positive effect on the success rate, but because most of these detection methods are in the research stage, the methodology recommended by the World Health Organization is too cumbersome and complicated, and the stability and repeatability of the methodology are poor, so it is difficult to be used as a routine clinical practice. The method is popularized, but there is a lack of industrialized products

Existing experimental methods require the entire test process to be completed in one go, from the pretreatment of semen samples, in vitro capacitation, calcium ion induction to fluorescent staining of smears and result interpretation, the entire test takes 6 to 7 hours, and the test is intensive and difficult to apply clinically and popularity

Moreover, the intensity of the fluorescent dyeing system of the existing test method is not high, and it usually decays rapidly within half an hour, which affects the accuracy of result interpretation and will cause instability of test results

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