CN109060781B - 血清中抗卵磷脂抗体检测方法 - Google Patents

血清中抗卵磷脂抗体检测方法 Download PDF

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CN109060781B
CN109060781B CN201810924002.8A CN201810924002A CN109060781B CN 109060781 B CN109060781 B CN 109060781B CN 201810924002 A CN201810924002 A CN 201810924002A CN 109060781 B CN109060781 B CN 109060781B
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李立和
王学超
牛建新
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Tianjin Baodi Hospital
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Abstract

本发明公开了一种血清中抗卵磷脂抗体测定方法，属于免疫分析技术领域，本发明技术路线为：以健康成年男性前列腺液中卵磷脂小体为基质制成的生物薄片，加入1：100PBS稀释的血清，20～37℃ 1～2h或4℃过夜，0.01mol/L PBS洗2min 3次；加入羊抗人IgG FITC荧光标记物，20～37℃ 1～2h，0.01mol/L PBS洗2min 4次；封片剂封片，荧光显微镜10ⅹ40倍视野下观察；FITC在490～495nm激发，在520～530nm发射荧光，如为卵磷脂小体呈黄绿色，即为血清中抗卵磷脂抗体阳性。

Description

血清中抗卵磷脂抗体检测方法
技术领域
本发明公开了一种血清中抗卵磷脂抗体检测方法，属于免疫学分析技术领域，通过测试反应的结果产生颜色变化来测试材料的方法。
背景技术
卵磷脂是人体每一个细胞不可缺少的一种物质，如果缺乏，就会造成降低皮肤细胞的再生能力，会导致皮肤粗糙，有皱纹，如果适当地摄取卵磷脂，那么皮肤就有良好的亲水性和亲油性，皮肤就会有光泽。卵磷脂当中所含的肌醇还是毛发的一种主要营养物，可以抑制脱发，使头发慢慢变黑，磷脂和蛋白质是构成细胞膜的一种主要成分，蛋黄当中还有丰富的卵磷脂，牛奶和动物的脑骨髓，心脏，肺部，肝脏，以及大豆和酵母中都含有卵磷脂，卵磷脂起着对血清脂质的调节作用，调节血清脂质水平，意味着可以降低胆固醇水平，保护肝脏也可以改善记忆力，加强免疫力以及抗脂肪肝的活力。在二十世纪六十年代的时候，科学家们就发现卵磷脂，具有保护心脏的作用，脑神经细胞当中的卵磷脂含量约占其含量的17％-20％，胆碱是大豆卵磷脂的基本成分，卵磷脂的充分供应会保证充分的胆碱与人体内的胰腺合成为乙酰胆碱，乙酰胆碱是大脑内的一种信息传导物质，从而会提高脑细胞的活性化程度，提高记忆力与智力水平。
卵磷脂是一种天然的解毒剂，它可以分解体内的毒素，通过增加血红素，为皮肤提供充分的水和氧气等营养成分会使皮肤变得光滑柔润，卵磷脂可以促进肠胃血液循环以及肠胃蠕动，还可以有助于预防以及改善便秘，如果有胆结石的患者每天坚持服用大豆卵磷脂，不仅可以防止胆结石形成，而且还可以不同程度地分解，消除已经形成的胆结石，长期饮酒或者是营养过剩以及脂肪肝的患者，每天服用10-15克的卵磷脂在3-5个月之后，血脂的指标可以恢复到正常水平。
发明内容：
为了解决现有技术中血清中抗卵磷脂抗体检测，本发明提供一种经济方便易行，准确性更高、通过免疫荧光技术检测血清中抗卵磷脂抗体的方法。
本发明的技术方案使以卵磷脂小体为反应基质抗原制成生物薄片，加入待检者血清，洗涤，加入羊抗人IgG荧光标记物，在荧光显微镜下观察，卵磷脂小体出现黄绿色荧光，即为阳性。
具体实施方式
1.生物薄片的制备取正常人前列腺液100μl，加入到含有30g/L N-乙酰-L-半胱氨酸的磷酸盐缓冲液进行洗涤5次，进行15000G离心10min，弃去上清液,用PBS调整卵磷脂小体数至100个/μl，点片到含有多聚赖氨酸玻片上,冷风吹干后,密封,-70℃低温保存。
2.抗卵磷脂抗体的检测取生物薄片放置室温后，加入1：100PBS稀释的血清，20～37℃1～2h或4℃过夜，0.01mol/L PBS洗2min 3次；加入羊抗人IgG FITC荧光标记物,20～37℃1～2h，0.01mol/L PBS洗2min 4次；封片剂封片，荧光显微镜10ⅹ40倍视野下观察；FITC在490～495nm激发，在520～530nm发射荧光，如为卵磷脂小体呈黄绿色，即为阳性。
其中，N-乙酰-L-半胱氨酸的巯基结构可使前列腺粘液蛋白的双硫键(-S-S-)断裂，降低粘稠度，使卵磷脂小体解离出来，易于点片。
3.为了更好地理解本发明，下面通过实验来进一步说明本发明在检测抗卵磷脂抗体方面的积极效果。
所选用的病例为2017年5月至2017年10月在我院神经内科住院患者86例，其中男性48例，平均年龄62.5岁；女性38例平均年龄62.0岁，早晨空腹采血2.0ml，进行抗卵磷脂抗体检测。
分别采用本发明方法和亚辉龙化学发光法进行血清中卵磷脂抗体检测，并将两种方法进行统计学分析。
表1两种方法检测结果比较
Figure GDF0000020066470000021
表2两种方法检测结果比较(％)
Figure GDF0000020066470000022
通过表1、表2可以看出本发明的测试方法在灵敏度,特异性,阳性预测值，阴性预测值等指标与化学发光法比较非常接近的，两种方法检测结果符合率大于80％，因此，可以应用本发明方法进行卵磷脂抗体的测定。

Claims (1)

1.一种血清中抗卵磷脂抗体检测方法，其特征在于以前列腺液中卵磷脂小体为基质制成的生物薄片，制备方法为：取正常人前列腺液100μl，加入到含有N-乙酰-半胱氨酸的磷酸盐缓冲液进行洗涤5次，进行15000G离心10min，弃去上清液， 用PBS调整卵磷脂小体数至100个/μl，点片到含有多聚赖氨酸玻片上,冷风吹干后,密封,-70℃低温保存。
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