中国血液净化

中国血液净化 ›› 2017, Vol. 16 ›› Issue (09): 607-611.doi: 10.3969/j.issn.1671-4091.2017.09.007

• 基础研究 • 上一篇    下一篇

超声辅助检测血浆内毒素的鲎试验方法及应用

李永桂1,杨正根1,何晓帆2,胡家亮1,陈校园1   

  1. 1. 广州康盛生物科技有限公司
    2. 国家食品药品监督管理局广州医疗器械质量监督检验中心
  • 收稿日期:2017-02-20 修回日期:2017-06-28 出版日期:2017-09-12 发布日期:2017-09-12
  • 通讯作者: 陈校园 chenxy@koncen.com E-mail:chenxy@koncen.com
  • 基金资助:

    广东省康盛血液净化吸附产品与技术的研发与应用院士工作站，编号：2016B090921006

The application of ultrasound pretreatment for the plasma endotoxin assay by limulus lysate test

  • Received:2017-02-20 Revised:2017-06-28 Online:2017-09-12 Published:2017-09-12

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摘要： 目的探讨超声对检测血浆内毒素的影响。方法以富血小板血浆和贫血小板血浆为研究对象，以超声等不同预处理血浆方式作为标准，分为6 组，A 组(超声辅助预处理富血小板血浆组)、B 组(无超声只漩涡振荡辅助预处理富血小板血浆组)、C 组(内毒素处理剂辅助预处理富血小板血浆组)、D 组(超声辅助预处理贫血小板血浆组)、E 组(无超声只漩涡振荡辅助预处理贫血小板血浆组)、F 组(内毒素处理剂辅助预处理贫血小板血浆组)，每组5 个样本，应用终点偶氮显色法鲎试剂定量检测血浆中微量内毒素。结果①A 组与B 组间回收率具有显著性差异(t=9.424，P＜0.001)，但最优回收率数量无显著性差异(c2=1.111，P=0.500)；A 组与C 组间回收率无显著性差异(t=-0.815，P=0.439)，且最优回收率数量也无显著性差异(c2=0.000，P=0.778)；D 与E 组间回收率具有显著性差异(t=30.395，P＜0.001)，但最优回收率数量无显著性差异(x2= 2.500, P=0.222)；D 与F 组间回收率无显著性差异(t=-0.375, P=0.717)，且最优回收率数量也无显著性差异(x2=0.000, P= 0.527)；A 与D 组间回收率无显著性差异(t=0.355，P=0.732)，且最优回收率数量也无显著性差异(x2=0.476，P=0.500)。② 超声法变异系数小，分别为2.7%，3.5%，且测试回收率在90%～100%之间，比国家标准要求的50%～200%更加准确。结论采用超声辅助预处理血浆检测方法具有成本低、灵敏度高、检测结果可靠和检测效率高的优点。预处理血浆检测方法具有成本低、灵敏度高、检测结果可靠和检测效率高的优点。

关键词: 内毒素, 超声法, 血浆, 检测

Abstract: Objective To explore the effect of ultrasound pretreatment for the detection of plasma endotoxin. Methods We used the platelet rich plasma and platelet poor plasma as the research objects. These plasma samples were pretreated with ultrasound or other methods and divided into 6 groups with 5 samples in every group: group A (platelet rich plasma pretreated with ultrasound), group B (platelet rich plasma pretreated with vortex), group C (platelet rich plasma pretreated with endotoxin reagent), group D (platelet poor plasma pretreated with ultrasound), group E (platelet poor plasma pretreated with vortex), and group F (platelet
poor plasma pretreated with endotoxin reagent). Trace endotoxin in plasma was assayed by limulus lysate reagent using end point azo chromogenic method. Results ①The recovery rate for endotoxin was significantly different, but the optimal recovery rate had no significant difference between group A and group B (t=
9.424, P＜0.001 for recovery rate; x2=1.111, P=0.500 for optimal recovery rate), and between group D and group E (t=30.395, P＜0.001 for recovery rate; x2=2.500, P=0.222 for optimal recovery rate). The recovery rate for endotoxin had no significant difference, so did the optimal recovery rate between group A and group C (t= - 0.815, P=0.439 for recovery rate; x2=0.000, P=0.778 for optimal recovery rate), between group D and group F (t= - 0.375, P=0.717 for recovery rate; x2=0.000, P=0.527 for optimal recovery rate), and between group A and group D (t=0.355, P=0.732 for recovery rate; x2=0.476, P=0.500 for optimal recovery rate). ② The variation coefficient of the results from the groups pretreated with ultrasound was narrower (2.7% and 3.5%), and the recovery rate was 90-100%, more accurate than the national standard of 50～200%. Conclusions The pretreatment with ultrasound for the detection of endotoxin in plasma has the advantages of lower cost, higher sensitivity, reliable result and higher efficiency.

Key words: endotoxin, ultrasonicmethod, plasma, detection