CAJ | 학술논문

目的：评价肠道病毒研究中常用的5种细胞对柯萨奇病毒A组16型（coxsackievirus A16， CA16）和肠道病毒71型（enterovirus 71，EV71）的敏感性，为分离和培养CA16和EV71提供实验室依据。方法用分离的各15株CA16和EV71分别感染 RD、Vero、KMB17、Hep2和L20B细胞，每天观察细胞病变情况，按Karber法计算病毒感染性滴度。采用不同的统计学方法（包括方差分析和卡方检验）比较5种细胞对CA16和EV71的敏感性差异。结果5种细胞的CA16和EV71病毒感染性滴度间的差异具有统计学意义（CA16：F＝18．481，P＝0．000；EV71：χ2＝63．106，P＝0．000）。在5种细胞中，RD细胞对2种病毒的敏感性最高，CA16和 EV71的病毒感染性滴度分别可达7．8和8．2 lgCCID50／ml ，且均于接种细胞后48 h达增殖高峰，第4天进入增殖平台期；2种病毒在 Hep2和L20B细胞中的病毒感染性滴度均较低，CA16分别为5．2和4．2 lgCCID50／ml ，EV71分别为4．9和4．0 lgCCID50／ml ，且2种病毒在接种后第6～7天才达增殖高峰，但仅L20B细胞中的2种病毒于接种后第7天进入增殖平台期；CA16在 Vero 和 KMB17细胞中的病毒感染性滴度分别为6．9和7．3 lgCCID50／ml ，EV71分别为7．1和6．8 lgCCID50／ml ，2种病毒均在接种Vero和KMB17细胞后第3天达到增殖高峰，第5天进入增殖平台期。结论 RD细胞对CA16和 EV71的敏感性最好，其次是Vero和KMB17细胞，而Hep2和L20B细胞并非是分离和培养这2种病毒的理想细胞。
목적：평개장도병독연구중상용적5충세포대가살기병독A조16형（coxsackievirus A16， CA16）화장도병독71형（enterovirus 71，EV71）적민감성，위분리화배양CA16화EV71제공실험실의거。방법용분리적각15주CA16화EV71분별감염 RD、Vero、KMB17、Hep2화L20B세포，매천관찰세포병변정황，안Karber법계산병독감염성적도。채용불동적통계학방법（포괄방차분석화잡방검험）비교5충세포대CA16화EV71적민감성차이。결과5충세포적CA16화EV71병독감염성적도간적차이구유통계학의의（CA16：F＝18．481，P＝0．000；EV71：χ2＝63．106，P＝0．000）。재5충세포중，RD세포대2충병독적민감성최고，CA16화 EV71적병독감염성적도분별가체7．8화8．2 lgCCID50／ml ，차균우접충세포후48 h체증식고봉，제4천진입증식평태기；2충병독재 Hep2화L20B세포중적병독감염성적도균교저，CA16분별위5．2화4．2 lgCCID50／ml ，EV71분별위4．9화4．0 lgCCID50／ml ，차2충병독재접충후제6～7천재체증식고봉，단부L20B세포중적2충병독우접충후제7천진입증식평태기；CA16재 Vero 화 KMB17세포중적병독감염성적도분별위6．9화7．3 lgCCID50／ml ，EV71분별위7．1화6．8 lgCCID50／ml ，2충병독균재접충Vero화KMB17세포후제3천체도증식고봉，제5천진입증식평태기。결론 RD세포대CA16화 EV71적민감성최호，기차시Vero화KMB17세포，이Hep2화L20B세포병비시분리화배양저2충병독적이상세포。
Objective To evalute sensitivities of five kinds of cells to coxsackievirus A16 (CA16) and Enterovirus 71 (EV71 ) and provide laboratory evidences for isolation and culture of 2 viruses . Methods RD ,Vero ,KMB17 ,Hep2 and L20B cells were infected with 15 CA16 and 15 EV71 clinical isolates ,respectively .The cytopathic effect of 5 cells were observed everyday .The infectious titers of CA16 and EV71 were calculated by the Karber method .The difference in sensitivity to CA16 and EV71 among the 5 cells were compared with statistical methods ,including one‐way ANOVA and chi‐square test .Results The infectious titers of CA16 and EV71 in 5 cells had significant difference (CA16 :F=18 .481 ,P=0 .000 ;EV71 :χ2 =63 .106 ,P=0 .000) .Among the 5 cells ,RD cell showed the highest sensitivity to 2 viruses , CA16 and EV71 , with infectious titers of 7 .8 and 8 .2 lgCCID50/ml , respectively .The proliferation of CA16 and EV71 reached peak at 48 h after inoculation and entered plateau phase 4 days after inoculation .The infectious titers of CA16 and EV71 in Hep2 and L20B were lower than those in other cells ,and the infectious titers in Hep2 and L20B cells were 5 .2 and 4 .2 lgCCID50/ml for CA16 and 4 .9 and 4 .0 lgCCID50/ml for EV71 ,respectively .The proliferation of CA16 and EV71 reached peak 6‐7 days after inoculation in L20B and Hep2 cells ,but entered plateau phase 7 days after inoculation only in L20B cell . The infectious titers in Vero and KMB17 cell were 6 .9 and 7 .3 lgCCID50/ml for CA16 and 7 .1 and 6 .8 lgCCID50/ml for EV71 ,respectively .The proliferation of of CA16 and EV71 reached peak 3 days after inoculation and entered plateau phase 5 days after inoculation . Conclusion Among five kinds of cells , RD cell has the best sensitivity to CA16 and EV71 , and sensitivities of Vero and KMB17 cells are next ,but Hep2 and L20B cells may not be ideal cells for isolation and culture of CA16 and EV71 .