Pediatric Gastroenterology, Hepatology & Nutrition

The Korean Society of Pediatric Gastroenterology, Hepatology and Nutrition (대한소아소화기영양학회)
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Usefulness of Escherichia coli-expressed Recombinant VP6 Proteins of Group A Rotavirus in Serodiagosis of Rotavirus Infection

대장균에서 발현된 A군 로타바이러스 VP6 단백질을 이용한 로타바이러스 감염의 혈청학적 진단의 유용성

  • Seo, Ji-Hyun (Department of Pediatrics, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Kim, So-Young (Department of Pediatrics, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Park, Ji-Sook (Department of Pediatrics, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Lim, Jae-Young (Department of Pediatrics, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Park, Chan-Hoo (Department of Pediatrics, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Woo, Hyang-Ok (Department of Pediatrics, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Youn, Hee-Shang (Department of Pediatrics, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Kim, Won-Yong (Department of Microbiology, Chung-Ang University College of Medicine) ;
  • Kang, Hyung-Lyun (Department of Microbiology, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Baik, Seung-Chul (Department of Microbiology, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Lee, Woo-Kon (Department of Microbiology, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Cho, Myung-Je (Department of Microbiology, Gyeongsang National University School of Medicine, Institute of Health Science) ;
  • Rhee, Kwang-Ho (Department of Microbiology, Gyeongsang National University School of Medicine, Institute of Health Science)
  • 서지현 (경상대학교 의학전문대학원 소아청소년과학교실) ;
  • 김소영 (경상대학교 의학전문대학원 소아청소년과학교실) ;
  • 박지숙 (경상대학교 의학전문대학원 소아청소년과학교실) ;
  • 임재영 (경상대학교 의학전문대학원 소아청소년과학교실) ;
  • 박찬후 (경상대학교 의학전문대학원 소아청소년과학교실) ;
  • 우향옥 (경상대학교 의학전문대학원 소아청소년과학교실) ;
  • 윤희상 (경상대학교 의학전문대학원 소아청소년과학교실) ;
  • 김원용 (중앙대학교 의과대학 미생물학교실) ;
  • 강형련 (경상대학교 의학전문대학원 미생물학교실, 건강과학연구원) ;
  • 백승철 (경상대학교 의학전문대학원 미생물학교실, 건강과학연구원) ;
  • 이우곤 (경상대학교 의학전문대학원 미생물학교실, 건강과학연구원) ;
  • 조명제 (경상대학교 의학전문대학원 미생물학교실, 건강과학연구원) ;
  • 이광호 (경상대학교 의학전문대학원 미생물학교실, 건강과학연구원)
  • Received : 2010.08.13
  • Accepted : 2010.09.03
  • Published : 2010.09.30
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Abstract

Purpose: The serologic diagnosis of rotaviral infections is not commonly used in clinical practice, but is used in seroepidemiologic studies. In this study, the usefulness of Escherichia coli-expressed recombinant VP6 proteins of group A rotavirus in the serodiagnosis of rotavirus infections by ELISA was evaluated. Methods: The recombinant VP6 proteins of group A rotavirus expressed in E. coli Rosetta II strain were purified and identified. One hundred sera from 22 children (4 healthy neonates, 13 healthy children, and 5 immunocompromised children) who had serial sera samples prior to and after rotavirus infections were provided by the Gyeongsang National University Hospital, a member of the National Biobank of Korea. IgG, IgA, and IgM antibodies against rVP6 were analyzed by ELISA in all of the patients and Western blot analysis in 4 neonates. Results: ELISA tests using rVP6 proteins of group A rotavirus as antigen revealed that IgG, IgA, and IgM antibodies increased after rotaviral infections in most neonates and healthy children. IgG antibodies also increased after rotaviral infections in most immunocompromised children without an adequate increase in IgM or IgA antibodies. Western blot analysis in four neonates revealed very early IgM antibody responses, even in the sera with low optical densities in ELISA tests. Conclusion: Our study showed that ELISA using rVP6 as an antigen is a valid diagnostic tool for seroepidemiologic studies of rotavirus infections and Western blot analysis is a sensitive test in detecting IgG, IgA, and and IgM antibodies in patients with rotavirus infections.

목 적: 로타바이러스 감염역학의 변화를 연구하기 위해 A군 로타바이러스의 VP6 유전자를 대장균에 발현시켜 확보한 rVP6 단백질이 항원성이 있는지를 확인하고 이것을 항원으로 한 효소면역측정법이 로타바이러스 IgG, IgA와 IgM 항체를 정량적으로 평가할 수 있는지 확인하고자 하였다. 방 법: 경상대학교병원에서 로타바이러스 감염을 진단받은 소아들 중 진단 받기 전, 진단 당시, 회복기 이후의 연속적인 혈청을 확보할 수 있었던 22명에게서 100개의 혈청을 경상대학교병원 인체자원은행으로부터 제공받아 로타바이러스 VP6 유전자를 클로닝 하여 대장균에 발현시켜 제조한 rVP6 항원으로 한 효소면역측정법으로 IgG, IgA와 IgM 항체 역가를 측정하였다. 이 중 건강한 신생아 4명에서는 면역 블로팅을 같이 시행하였다. 결 과: 건강한 신생아와 영유아 17명에서 감염 후 확보된 혈청에서 IgG, IgA, IgM 항체 중 최소한 한 종류의 항체 역가 증가가 동반되어 있었다. 면역이 저하된 소아 5명 중 4명에서는 IgG 항체 역가는 증가되었으나 IgA 항체 역가는 2명에서만 증가하였고, IgM 항체 역가는 5명 모두 증가하지 않았다. 신생아 4명에서 시행된 면역 블로팅 검사에서는 IgM 항체인 경우는 효소면역측정법보다 예민하게 진단 초기부터 4명 모두 양성으로 판정되었다. 결 론: A군 로타바이러스의 VP6 유전자를 대장균에 발현시켜 확보한 rVP6 단백질은 항원성이 있으며 이것을 항원으로 한 효소면역측정법은 로타바이러스 감염후 IgG, IgA, IgM 항체 역가 증가를 정량적으로 평가할 수 있어 지역사회에서 발생한 로타바이러스 감염역학의 변화를 연구하는데 유용할 것으로 판단된다.

Keywords

References

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