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수두바이러스의 정량에 있어서 Solid-overlay Standard Plaque Assay와 Immunofocus Assay의 효용성 비교 연구
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  • 수두바이러스의 정량에 있어서 Solid-overlay Standard Plaque Assay와 Immunofocus Assay의 효용성 비교 연구
  • Comparative Study of the Standard Plaque Assay with Solid-overlay and Immunofocus Assay for Varicella-zoster Virus Titration
저자명
이화경,정용석,Lee. Hwa-Kyung,Jeong. Yong-Seok
간행물명
대한바이러스학회지
권/호정보
2000년|30권 1호|pp.61-70 (10 pages)
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대한바이러스학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Standard plaque assay using agarose-overlay has long been used for titration of many infectious virus particle. Plaque assay for the titration of varicella-zoster virus and its live vaccine requires three intermittent agarose overlay to visualize plaques. Overall procedure of the assay takes at least nine days from virus inoculation and microbe contamination including fungi is frequently accompanied during incubation period. We studied whether an immunofocus assay in conjunction with peroxidase-mediated immunohistochemical reaction may replace the standard plaque assay for the virus titration by comparing the two methods. A linear relationship was observed between number of foci and virus dilution. The number of foci in a given dilution of virus appeared a little higher than counted plaques formed in standard plaque assay. Independent titration results obtained from two assay methods for a given dilution of virus demonstrated a strong correlation ($r^2=0.99$). Foci of virus infected cells as revealed by the enzyme reaction could be counted either 4 days post-infection (p.i.) under low magnification (40X) microscopy, or 6 days p.i. by naked eye observation. Larger size of cell cuture plate, virus adsorption at $35^{circ}C$, and 10% FBS in diluent appeared to be better conditions for the assay. Immunofocus assay will be an effective and dependable titration method for varicella-zoster virus and its live vaccine in place of the standard plaque assay in respect to accuracy, costs, and experimental convenience.