AMPLIFICATION AND DETECTION OF LENTIVIRAL DNA INSIDE CELLS
AMPLIFICATION AND DETECTION OF LENTIVIRAL DNA INSIDE CELLS
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DOI：
10.1073/pnas.87.13.4971
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发表时间：
1990-07-01
影响因子：
11.1
通讯作者：
STASKUS, KA
中科院分区：
文献类型：
--
作者：
HAASE, AT;RETZEL, EF;STASKUS, KA
维斯纳病毒和人类免疫缺陷病毒分别是动物和人类慢病毒的原型，尽管宿主免疫应答，但它们仍然存在并传播，因为组织和血流中的细胞以隐蔽状态携带病毒基因组。为了便于识别这些潜伏感染的细胞，聚合酶链反应已被调整为扩增固定细胞中的病毒DNA，以通过原位杂交进行检测。通过使用产生具有重叠粘性末端的DNA片段的多引物组，可以在感染的细胞中扩增、保留和检测维斯纳病毒DNA，其灵敏度超过现有方法超过2个数量级。单细胞技术的这一进展应证明有助于诊断和深入了解病毒感染的发病机制，并为寻找病因不明的慢性疾病中的病毒提供新的机会。
Visna virus and human immunodeficiency virus are prototypes of animal and human lentiviruses, respectively, that persist and are disseminated despite the host immune response because cells in the tissues and the blood stream harbor viral genomes in a covert state. To facilitate identification of these latently infected cells, the polymerase chain reaction has been adapted to amplify viral DNA in fixed cells for detection by in situ hybridization. By using a multiple primer set that generates DNA segments with overlapping cohesive termini, visna virus DNA can be amplified, retained, and detected in infected cells with sensitivities that exceed those of existing methods by more than 2 orders of magnitude. This advance in single-cell technology should prove useful in diagnosing and gaining insight into the pathogenesis of viral infections and provide new opportunities to look for viruses in chronic diseases of unknown etiology.