Tuberculosis and Respiratory Diseases

  • 제60권4호
  • /
  • Pages.412-418
  • /
  • 2006
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  • 1738-3536(pISSN)
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  • 2005-6184(eISSN)
대한결핵및호흡기학회 (The Korean Academy of Tuberculosis and Respiratory Diseases)
권호 표지

단백질 칩을 이용한 클라미디아 폐렴의 진단

Development of Protein Chip for Diagnosis of Chlamydophia Pneumoniae

  • 김우진 (강원대학교 의과대학 내과학 교실) ;
  • 이희영 (강원대학교 의과대학 내과학 교실) ;
  • 이승준 (강원대학교 의과대학 내과학 교실) ;
  • 정세희 (강원대학교 의과대학 생화학교실) ;
  • 육종설 (강원대학교 의과대학 생화학교실) ;
  • 하권수 (강원대학교 의과대학 생화학교실) ;
  • 정기석 (한림대학교 의과대학 내과학교실)
  • Kim, Woo Jin (Department of Internal Medicine, College of Medicine, Kangwon National University) ;
  • Lee, Hui Young (Department of Internal Medicine, College of Medicine, Kangwon National University) ;
  • Lee, Seung-Joon (Department of Internal Medicine, College of Medicine, Kangwon National University) ;
  • Jung, Se-Hui (Department of Molecular and Cellular Biochemistry, College of Medicine, Kangwon National University) ;
  • Yuk, Jong Seol (Department of Molecular and Cellular Biochemistry, College of Medicine, Kangwon National University) ;
  • Ha, Kwon-Soo (Department of Molecular and Cellular Biochemistry, College of Medicine, Kangwon National University) ;
  • Jung, Ki-Suck (Department of Internal Medicine, College of Medicine, Hallym University)
  • 투고 : 2006.03.10
  • 심사 : 2006.04.17
  • 발행 : 2006.04.30
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초록

연구배경 : 클라미디아 감염의 진단은 혈청검사로 이루어진다. 현재 표준 방법은 MIF(microimmunofluorescence)이나 이 방법은 주관적이고 시간이 많이 걸리는 단점이 있다. 최근을 SPR(surface plasmon resonance) 센서를 이용한 단백질 칩이 감염의 새로운 진단 방법으로 제시되고 있다. 클라미디아 감염의 진단을 위한 단백질 칩 개발을 위하여 금 칩 표면에 세균을 고정하고 클라미디아 균에 대한 항체와 표면 위 세균과의 반응을 SPR 센서를 이용하여 측정하고자 하였다. 방법 : 표면 항원으로 배양한 Chlamydophila pneumoniae LKK1의 EB를 정제하였다. 양전하를 띤 PDDA (polydiallyldimethylammonium chloride)를 이용하여 전하를 이용한 단백질 칩을 제작하였다. 클라미디아 균을 고정시킨 후에 atomic force microscopy를 이용하여 표면을 관찰하였다. 클라미디아 균에 대한 항체를 투여하고 나서 자체 제작한 SPR 센서를 이용하여 항원 항체 반응을 SPR 파장 변화로 측정하였다. 결과 : 양전하를 띤 PDDA 표면위에서 클라미디아 균이 고정되었음을 확인 하였다. 그리고, 항체를 투여한 후에 SPR 파장의 증가를 확인하였다. 파장 변화는 항원의 농도와 관련이 있었다. 결론 : 전하를 이용하여 클라미디아 폐렴균의 EB를 단백질 칩에 고정하였고, 단백질 칩 위에서의 항원 항체 반응을 확인하였다. 비정형 폐렴의 진단에 SPR 센서가 기여할 수 있을 것으로 사료되나, 실제 임상 시료에의 적용을 위해서는 좀더 연구가 필요할 것으로 사료된다.

Background; The diagnosis of chlamydial infection is based on serology. The current gold standard of diagnosis is MIF(microimmunofluorescence), but this modality is subjective and time-consuming. Protein microarray with using a SPR(surface plasmon resonance) sensor has recently been suggested as a method for detecting infection. For developing a protein chip to diagnose chlamydial infection, EBs(elementary bodies) were immobilized on a gold chip and the interaction between an antibody for Chlamydophila pneumoniae and the EBs(elementary bodies) immobilized on the surface of the gold chip was measured by using an SPR sensor. Methods; For the surface antigen, the EBs of Chlamydophila pneumoniae LKK1 were purified. Charged arrays were prepared by using PDDA(polydiallyldimethylammonium chloride) which has a positive charge. After immobilization of the chlamydial EBs on the PDDA surface, the investigation of the surface was done with using atomic force microscopy. After the antibody for C. pneumoniae was applied on chip, we monitored the SPR wavelength-shift to detect any antigen-antibody interaction with using a self-assembled SPR sensor. Results; The chlamydial EBs on the positively charged PDDA were visible on the surface with using atomic force microscopy. The SPR wavelength increased after interaction of antibody for C. pneumoniae with the EBs immobilized on charged gold surface. The wavelength-shift was correlated with the concentration of antigens. Conclusion; The surface immobilization of EBs on the gold surface with the charged arrays was identified and the antigen-antibody interaction on the gold chip was detected via the SPR sensor. Further investigations are needed to apply this technique to the clinical field.

키워드

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