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Comparison of Three Methods Used to Isolate Dengue Virus Type 2

机译：三种分离2型登革热病毒的方法的比较

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During the 1969 dengue epidemic in Puerto Rico, human sera and Aedes aegypti mosquitoes were collected for virus isolation and identification. Three methods of isolation were used and compared. In the first method, we inoculated newborn mice by the intracranial route, noted any signs of illness, and serially passed specimens in mice until virus was isolated. In the second method, we inoculated tube cultures of LLC-MK2 cells, noted any cytopathic effect (CPE), and assayed fluids for virus by plaque formation in LLC-MK2 cell monolayers. The third method was different from the second only in that the original specimens were first inoculated into fluid cultures of Singh's A. albopictus cells. No significant CPE was seen in LLC-MK2 cultures; however, distinct syncytial CPE was observed in A. albopictus cells. About the same number of virus isolates were made in each isolation system. Virus isolates from both sera and mosquitoes were identified as dengue type 2 by a plaque-reduction neutralization test in LLC-MK2 cells. The utility of the three methods, individually or in combination, is discussed and related to diagnostic and epidemic situations.
机译：在1969年波多黎各的登革热流行期间，收集了人的血清和埃及伊蚊，以进行病毒分离和鉴定。使用了三种隔离方法并进行了比较。在第一种方法中，我们通过颅内途径接种了新生小鼠，注意到有任何疾病迹象，并在小鼠中连续通过标本直到分离出病毒。在第二种方法中，我们接种了LLC-MK2细胞的试管培养物，注意到了任何细胞病变作用（CPE），并通过在LLC-MK2细胞单层中形成噬菌斑来分析液体中的病毒。第三种方法与第二种方法的不同之处仅在于首先将原始标本接种到Singh's albopictus细胞的液体培养物中。在LLC-MK2培养物中未观察到明显的CPE。然而，在白纹曲霉细胞中观察到了明显的合胞体CPE。在每个隔离系统中制作了大约相同数量的病毒隔离株。通过LLC-MK2细胞中的噬斑减少中和测试，从血清和蚊子中分离出的病毒被鉴定为登革热2型。讨论了这三种方法的实用性，无论是单独使用还是组合使用，都与诊断和流行情况有关。

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