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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Diagnosis of human immunodeficiency virus infection using citrated whole blood.
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Diagnosis of human immunodeficiency virus infection using citrated whole blood.

机译：使用柠檬酸盐全血诊断人类免疫缺陷病毒感染。

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Standard isolation of human immunodeficiency virus type 1 (HIV-1) from peripheral blood mononuclear cells (PBMC) requires 5 to 20 ml of blood, and the centrifugal separation of PBMC is expensive and time-consuming. Whole-blood coculture techniques use small sample volumes, do not require centrifugation, and allow measurement of the total viral burden in peripheral circulation. We compared the results of citrated whole-blood coculture with those obtained by the standard AIDS Clinical Trials Group PBMC semiquantitative culture method and reverse transcription-PCR quantitation of plasma HIV-1 RNA levels. PBMC cocultures were also set up with added erythrocytes (RBCs) to determine if the presence of RBCs affects the replication of HIV-1 in vitro. The mean number of cells required for a p24-positive PBMC coculture was approximately seven times greater than that required for a positive citrated whole-blood coculture (P < 0.01). At volumes of 100, 50, and 25 microl, the sensitivities of the whole-blood coculture were 94.5, 93.6, and 87.3%, respectively. The PBMC culture in the presence of added RBCs was more sensitive than PBMC coculture alone. The citrated whole-blood coculture was simple to perform, produced a reliable diagnosis of HIV infection in adult volunteers, was more sensitive than previously reported techniques even in half the culture time, and showed less variability than the PBMC coculture. Citrated whole-blood coculture may be a useful and efficient tool for diagnosing infection with HIV-1.
机译：从外周血单核细胞（PBMC）中标准分离1型人类免疫缺陷病毒（HIV-1）需要5到20 ml血液，而PBMC的离心分离既昂贵又费时。全血共培养技术使用的样品量很小，不需要离心，并且可以测量外周循环中的总病毒负荷。我们将柠檬酸盐全血共培养的结果与标准AIDS临床试验小组PBMC半定量培养方法和血浆HIV-1 RNA水平的反转录PCR定量结果进行了比较。 PBMC共培养物中还添加了红细胞（RBC），以确定RBC的存在是否影响HIV-1的体外复制。 p24阳性PBMC共培养所需的平均细胞数约为阳性柠檬酸盐全血共培养所需的平均细胞数的7倍（P <0.01）。在100、50和25微升的体积下，全血共培养的敏感性分别为94.5％，93.6％和87.3％。在存在添加的RBC的情况下，PBMC培养比单独的PBMC共培养更为敏感。柠檬酸全血共培养操作简单，可对成年志愿者的HIV感染进行可靠的诊断，比以前报道的技术甚至在一半的培养时间内都更加敏感，并且变异性比PBMC共培养小。柠檬酸全血共培养可能是诊断HIV-1感染的有用且有效的工具。

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