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Manufacturing and applications of human sperm hyaluronidase kit

A technology of hyaluronidase and sodium hyaluronate, which is applied in the preparation of test samples, measuring devices, and material analysis through optical means, can solve the problems of high cost and complicated operation, and achieve a vast territory and promising application prospects Good results

Inactive Publication Date: 2018-03-16
高晓勤 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] To sum up, the problems existing in the prior art are: the current method for detecting hyaluronidase in sperm is complex in operation and high in cost.

Method used

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  • Manufacturing and applications of human sperm hyaluronidase kit
  • Manufacturing and applications of human sperm hyaluronidase kit
  • Manufacturing and applications of human sperm hyaluronidase kit

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Embodiment Construction

[0024] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0025] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0026] Such as figure 1 As shown, the preparation method of the human pure hyaluronidase kit provided by the embodiments of the present invention comprises the following steps:

[0027] S101: Fabrication and fixation of the substrate film;

[0028] S102: After the collected fresh semen sample is liquefied, put it into a 5ml centrifuge tube and centrifuge at a relative centrifugal force of 864×g for 10min, discard the seminal plasma, add 2ml of phosphate buffer saline (PBS) with a pH ...

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Abstract

The invention belongs to the technical field of sperm hyaluronidase measurement, and discloses manufacturing and applications of a human sperm hyaluronidase kit. Sodium hyaluronate and a gelatin mixedsolution are used to prepare a sol-like matrix solution, then the sol-like matrix solution is painted and fixed to form a uniform base membrane for display activity of HYD; then sperms are incubatedon the ultrathin sodium hyaluronate-gelatin base membrane, centrifugation is performed, a phosphate buffer solution is used to wash the membrane inhibition factors to depolymerize sperm acrosome; thesodium hyaluronate-gelatin base membrane is released and dissolved, a pore area that surrounds the front part of sperms is formed; the sperms are observed by a light microscope, light penetrates the pore area, bright aureole appears on the periphery of the sperm heads; after dyeing, the enzyme reactions can be observed under a dark purplish blue background, and the HYD activity of each sperm can be estimated by measuring the diameter of each bright ring. The method is simple and the cost is low. The positive reaction rate of HYD can be calculated accurately. Stronger sperms are screened out for the test-tube baby technology, and the success rate is improved.

Description

technical field [0001] The invention belongs to the technical field of sperm hyaluronidase measurement, in particular to the manufacture and application of a human sperm hyaluronidase kit. Background technique [0002] Human sperm acrosome contains 24 enzymes known to humans, among which hyaluronidase (HYD) and acrosin are most closely related to fertilization. HYD plays an important role in the union of sperm and egg during fertilization. Determination of sperm HYD activity is one of the indicators to evaluate sperm function. In the previous research methods on sperm acrosome enzymes, most of them used biochemical methods to detect the enzyme activity, positive rate and enzyme reaction time of all sperm in the whole sample. For example, the determination of sperm acrosome arginine amidase activity requires expensive reagents such as polyethylene glycol octylphenyl ether (Triton solution), benzoyl arginyl p-nitroanilide (BAPNA solution), and can only react The acrosinase ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N21/84
CPCG01N1/30G01N21/84
Inventor 高晓勤杨喆王珂戴研平杨燕平马晓萍秦海霞倪佳杨建辉李淑钗
Owner 高晓勤
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