Montagne P, Cuillière M L, Molé C, Béné M C, Faure G
GRIP, Immunology Laboratory, Faculty of Medicine, Vandoeuvre les Nancy, France.
Clin Chem. 1998 Aug;44(8 Pt 1):1610-5.
Quantitation of lysozyme in human milk was performed by a microparticle-enhanced nephelometric immunoassay based on the measurement of the light scattered during the competitive immunoagglutination of a microparticle-lysozyme conjugate with an anti-lysozyme antiserum. This immunoassay has a detection limit of 8 microg/L of reaction mixture and can be performed using diluted milk (1:6000, in reaction mixture), excluding sample pretreatment. Human milk lysozyme can be quantified over the concentration range 0.09-1.50 g/L, with within- and between-run coefficients of variation <5%. Changes in the lysozyme concentration of human milk during lactation were determined in 636 samples. Lysozyme concentrations (mean +/- SE) decreased from colostrum (0.36 +/- 0.02 g/L) to transitional milk (0.30 +/- 0.01 g/L) and mature milk during days 15-42 (0.30 +/- 0.01 g/L), then increased in the mature milk during days 43-56 (0.35 +/- 0.01 g/L) and especially during days 57-84 (0.83 +/- 0.05 g/L). The proportion of lysozyme contributing to total protein was found to rise during lactation and was as follows: colostrum (1.7%), transitional milk (2.3%), and mature milk from days 15-28 (2.7%), days 29-42 (3.1%), days 43-56 (3.8%), and days 57-84 (7.3%). The assay developed for milk was also suitable for the determination of lysozyme in other human body fluids.
采用基于微粒 - 溶菌酶结合物与抗溶菌酶抗血清竞争性免疫凝集过程中光散射测量的微粒增强比浊免疫分析法对人乳中的溶菌酶进行定量。该免疫分析法对反应混合物的检测限为8μg/L，可使用稀释后的乳汁（在反应混合物中为1:6000）进行检测，无需样品预处理。人乳溶菌酶在0.09 - 1.50 g/L的浓度范围内可进行定量分析，批内和批间变异系数均<5%。对636份样品测定了哺乳期人乳中溶菌酶浓度的变化。溶菌酶浓度（均值±标准误）从初乳（0.36±0.02 g/L）降至过渡乳（0.30±0.01 g/L）以及第15 - 42天的成熟乳（0.30±0.01 g/L），然后在第43 - 56天的成熟乳中升高（0.35±0.01 g/L），尤其在第57 - 84天升高明显（0.83±0.05 g/L）。发现溶菌酶占总蛋白的比例在哺乳期上升，具体如下：初乳（1.7%）、过渡乳（2.3%）、第15 - 28天的成熟乳（2.7%）、第29 - 42天的成熟乳（3.1%）、第43 - 56天的成熟乳（3.8%）以及第57 - 84天的成熟乳（7.3%）。所开发的乳汁检测方法也适用于测定其他人体体液中的溶菌酶。
