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内毒素测量:气溶胶采样及一种新型鲎试剂法的应用

Endotoxin measurement: aerosol sampling and application of a new Limulus method.

作者信息

Milton D K, Gere R J, Feldman H A, Greaves I A

机构信息

Department of Environmental Science and Physiology, Harvard School of Public Health, Boston, MA 02115.

出版信息

Am Ind Hyg Assoc J. 1990 Jun;51(6):331-7. doi: 10.1080/15298669091369754.

Abstract

A new Limulus test was designed specifically for measuring endotoxin in environmental aerosols. The new Limulus method has a detection limit for airborne endotoxin of 2.2 pg (NP-1 activity)/m3 and can precisely quantitate aerosols containing 14 or more pg/m3 from samples representing 6.5 m3 of air. Aerosols in the range 100 to 500 pg/m3 were measured with 95% confidence of +/- 32% and an aerosol of 10 pg/m3 with 95% confidence of +/- 50%. Qualitative information about airborne endotoxin was also obtained from the assay. A wide variety of filter media were found to inactivate lipopolysaccharide in solution. This implies that airborne endotoxin can be measured only relative to the conditions of a particular study and that comparison of endotoxin aerosol measurements made under different circumstances are invalid. The research and policy implications of this observation are discussed. The need for improved collection and extraction methods notwithstanding, it was proposed that a buffered, parallel-line Limulus assay method be adopted as the standard method for measuring environmental endotoxin. The kinetic-turbidimetric Limulus assay with resistant-parallel-line estimates (KLARE), rate response method, should be considered a prime candidate for the standard method because of its precision, sensitivity, resistance to interference by pH, internal validation of estimates, and ability to provide qualitative as well as quantitative information about airborne endotoxin.

摘要

一种专门设计用于测量环境气溶胶中内毒素的新型鲎试剂检测法。这种新型鲎试剂检测法对空气中内毒素的检测限为2.2皮克(NP - 1活性)/立方米,并且能够精确地对来自代表6.5立方米空气的样本中含量为14皮克/立方米及以上的气溶胶进行定量分析。对于100至500皮克/立方米范围内的气溶胶,测量的95%置信区间为±32%;对于10皮克/立方米的气溶胶,测量的95%置信区间为±50%。通过该检测法还获得了有关空气中内毒素的定性信息。发现多种过滤介质可使溶液中的脂多糖失活。这意味着只能相对于特定研究的条件来测量空气中的内毒素,并且在不同情况下进行的内毒素气溶胶测量结果的比较是无效的。讨论了这一观察结果对研究和政策的影响。尽管需要改进收集和提取方法,但仍建议采用缓冲平行线鲎试剂检测法作为测量环境内毒素的标准方法。具有抗性平行线估计的动力学比浊法鲎试剂检测法(KLARE),即速率响应法,因其精确性、灵敏度、抗pH干扰性、估计值的内部验证以及能够提供有关空气中内毒素的定性和定量信息,应被视为标准方法的主要候选方法。

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