NEW TECHNIQUE FOR ASSAY OF INFECTIVITY OF HUMAN ADENOVIRUS 5 DNA
NEW TECHNIQUE FOR ASSAY OF INFECTIVITY OF HUMAN ADENOVIRUS 5 DNA
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DOI：
10.1016/0042-6822(73)90341-3
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发表时间：
1973-01-01
期刊：
影响因子：
3.7
通讯作者：
VANDEREB, AJ
中科院分区：
文献类型：
--
作者：
GRAHAM, FL;VANDEREB, AJ
建立了一种检测腺病毒5型DNA感染性的新方法.将病毒DNA稀释在含有低浓度磷酸盐的等渗盐水中，并加入氯化钙，从而形成磷酸钙沉淀。DNA与磷酸钙共沉淀，当将所得悬浮液加入人KB细胞单层时，DNA被吸附到细胞上。吸附后，在持续存在额外氯化钙的情况下，在37 ° C的液体培养基中孵育期间，DNA被吸收到细胞中。对于腺病毒5 DNA，该测定导致的噬斑比使用DEAE-葡聚糖获得的噬斑多100倍。此外，证明了每个培养物接种的DNA量与产生的噬菌斑数量之间的可重复关系。在高浓度（10-30 μg/ml）时，该检测方法最有效;低于此范围时，需要加入载体DNA以获得最佳结果。除腺病毒5 DNA外，该技术已成功用于检测腺病毒1和猿猴病毒40的DNA感染性。
A new technique for assaying infectivity of adenovirus 5 DNA has been developed. Viral DNA was diluted in isotonic saline containing phosphate at a low concentration, and calcium chloride was added, resulting in the formation of a calcium phosphate precipitate. DNA coprecipitated with the calcium phosphate and, when the resulting suspension was added to human KB cell monolayers, became adsorbed to the cells. Following adsorption, uptake of DNA into the cells occurred during an incubation in liquid medium at 37 ° in the continued presence of extra calcium chloride.For adenovirus 5 DNA the assay resulted in up to 100-fold more plaques than could be obtained using DEAE-dextran. Furthermore a reproducible relationship between amounts of DNA inoculated per culture and numbers of plaques produced was demonstrated. The assay was most efficient at high DNA concentrations (10–30 μg/ml); below this range the addition of carrier DNA was necessary for optimum results.In addition to adenovirus 5 DNA, the technique has been used successfully to assay infectivity of DNA from adenovirus 1 and simian virus 40.