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Development of a rapid, sensitive, and reproducible laboratory test kit for the assessment of plasma membrane integrity of human sperm.

Abstract：

OBJECTIVE:To develop a rapid, sensitive, and reproducible hypoosmotic swelling test kit for the assessment of plasma membrane integrity of human sperm in vitro. DESIGN:Prospective comparison of results with the World Health Organization (WHO) method, performed simultaneously. SETTING:Infertility center in a major city in India. PATIENT(S):Couples who presented for infertility evaluation. INTERVENTION(S):The sperm tail-coiling pattern representing sperm plasma membrane integrity was analyzed by using different concentrations of NaCl and also with plain double distilled water (ddH(2)O). Hypoosmotic swelling solution with 2% NaCl in ddH(2)O equivalent to 68 Osm/L was selected for further analysis because it provided the highest and the qualitatively best type (g) of tail coiling among the various other options tried in comparison. MAIN OUTCOME MEASURE(S):A rapid laboratory test kit for the assessment of plasma membrane integrity of sperm was developed that is equally sensitive and reproducible as that described in the WHO protocol. The test was validated by using different normal and subnormal semen samples and in comparison with the standard WHO protocol. RESULT(S):After the screening with different hypoosmotic solutions by using semen samples from fertile volunteers, NaCl (2%) provided the highest and the best types of typical tail coiling, characteristic of sperm with good plasma membrane integrity. The study was then extended to 60 normozoospermic semen samples, which demonstrated a hypoosmotic swelling response of 69.5% +/- 5.23% and 71.5% +/- 4.89%, as per WHO or the present modified method, respectively (coefficient of correlation, r = 0.741). In 13 oligospermic and 18 teratozoospermic subjects, the hypoosmotic swelling response of sperm as per the new method were 36.4% +/- 5.75% and 34.2% +/- 7.78%, respectively, which were comparable to those obtained through the WHO method. There was a 15% drop in viability of sperm after the hypoosmotic challenge (5 min), irrespective of the hypoosmotic solution used. No other morphological alterations in sperm were observed after the hypoosmotic challenge. CONCLUSION(S):The new test kit can be used routinely in laboratories for assessment of plasma membrane integrity of sperm in vitro.

journal_name

Fertil Steril

journal_title

Fertility and sterility

authors

Misro MM,Chaki SP

doi

10.1016/j.fertnstert.2007.02.003

subject

Has Abstract

pub_date

2008-01-01 00:00:00

pages

223-7

issue

eissn

0015-0282

issn

1556-5653

pii

S0015-0282(07)00341-X

journal_volume

pub_type

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Development of a rapid, sensitive, and reproducible laboratory test kit for the assessment of plasma membrane integrity of human sperm.