Gubler D J, Kuno G, Sather G E, Velez M, Oliver A
Am J Trop Med Hyg. 1984 Jan;33(1):158-65. doi: 10.4269/ajtmh.1984.33.158.
During the fall of 1981, a new method for the routine isolation and identification of dengue viruses in Puerto Rico was implemented utilizing C6/36 cell cultures and serotype specific antidengue monoclonal antibodies. A blind comparison of the monoclonal antibody indirect fluorescent antibody test (IFAT) and the complement fixation (CF) test for identification of 89 newly isolated dengue viruses of all four serotypes from the Caribbean, Asia and Africa showed 100% agreement. Although virus isolation rates were slightly lower than with the mosquito inoculation technique, use of the C6/36 cell culture system was much less time-consuming and allowed the processing of larger numbers of sera. Beginning in November 1981, a new virologic surveillance system was begun in Puerto Rico. Acute sera from persons with suspected dengue were selected for virus isolation attempts on the basis of geographic area of residence on the island, day after onset the blood was taken and clinical signs and symptoms. These sera were processed for virus isolation in C6/36 cell cultures, and virus isolates were identified by the IFAT using the monoclonal antibodies. Using this system, 2,702 sera were tested from November 1981 through August 1982. Dengue virus was isolated from 518, for an isolation rate of 19.2%. Dengue 1 was the predominant virus until December 1981, when dengue 4 became dominant. The changing patterns of dengue 1 and 4 distribution by time and geographic location on Puerto Rico were followed. This system allows the dengue viruses being transmitted in an area to be monitored with a minimal amount of effort and provides the early warning capability necessary to predict epidemic dengue.
1981年秋季,波多黎各实施了一种利用C6/36细胞培养物和血清型特异性抗登革热单克隆抗体对登革热病毒进行常规分离和鉴定的新方法。对单克隆抗体间接荧光抗体试验(IFAT)和补体结合试验(CF)进行盲法比较,以鉴定来自加勒比地区、亚洲和非洲的89株新分离的所有四种血清型的登革热病毒,结果显示二者符合率达100%。虽然病毒分离率略低于蚊虫接种技术,但使用C6/36细胞培养系统耗时少得多,且能处理更多血清样本。从1981年11月开始,波多黎各启动了一个新的病毒学监测系统。根据岛上居民的居住地理区域、采血日期以及临床症状和体征,选择疑似登革热患者的急性期血清进行病毒分离尝试。这些血清样本在C6/36细胞培养物中进行病毒分离处理,分离出的病毒用单克隆抗体通过IFAT进行鉴定。利用该系统,1981年11月至1982年8月期间共检测了2702份血清样本。从其中518份样本中分离出了登革热病毒,分离率为19.2%。在1981年12月之前,登革热1型是主要病毒,之后登革热4型成为优势病毒。对波多黎各登革热1型和4型在时间和地理位置上的分布变化模式进行了跟踪。该系统能够以最少的工作量监测某地区传播的登革热病毒,并提供预测登革热流行所需的早期预警能力。