Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Development of an Improved Endotoxin Detection Method Using Centrifugation

원심분리법을 이용한 혈청 내 내독소의 개선된 측정방법 연구

  • Choi, Hyeong Jwa (Department of Biomedical Science & Technology, Institute of Biomedical Science & Technology (IBST), Konkuk University) ;
  • Lim, Yoo Jung (Department of Biomedical Science & Technology, Institute of Biomedical Science & Technology (IBST), Konkuk University) ;
  • Lee, Eun Hee (Diatech Korea Co., LTD.) ;
  • Park, Jin Yeon (Department of Biomedical Science & Technology, Institute of Biomedical Science & Technology (IBST), Konkuk University) ;
  • Prabagar, Miglena G. (Department of Biomedical Science & Technology, Institute of Biomedical Science & Technology (IBST), Konkuk University) ;
  • Park, Hyung Soon (Diatech Korea Co., LTD.) ;
  • Kang, Young Sun (Department of Biomedical Science & Technology, Institute of Biomedical Science & Technology (IBST), Konkuk University)
  • Received : 2013.02.01
  • Accepted : 2013.04.11
  • Published : 2013.06.28
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Abstract

Endotoxins are part of the outer membrane of the cell wall of gram-negative bacteria and are continuously released during bacterial growth. Endotoxins typically induce severe sepsis and septic shock, which cause more than 50% of mortalities. Endotoxins are easily measured in the serum by the limulus amebocyte lysate (LAL) test. However, a nonspecific result is obtained, because the high concentration of serum proteins disturbs the enzyme reaction of the LAL test. In order to solve this problem, the LAL test was performed in this study after the centrifugation of the boiled serum samples to remove the impurities. As a result, among the various conditions examined, endotoxin measurement with the LAL test was the most accurate and repeatable after centrifugation of the boiled serum at $100^{\circ}C$. Moreover, the endotoxin was accurately and repeatedly measured from the prepared sera of mice that had been administered an intraperitoneal injection of purified lipopolysaccharides (LPS) or E. coli. Therefore, the application of centrifugation to remove impurities from boiled serum gives an accurate measurement of endotoxins in the sera of normal subjects or patients, and this will lead to the improved diagnosis and prevention of diseases caused by endotoxins. In addition, the centrifugation of boiled serum samples should be considered and included in the development of endotoxin test kits.

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