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用于检测脐带血捐献者中人疱疹病毒7型感染的定量聚合酶链反应

Quantitative polymerase chain reaction for detection of human herpesvirus-7 infection in umbilical cord blood donors.

作者信息

Abedi E, Kheirandish M, Sharifi Z, Samiee S, Kokhaei P, Pourpak Z, Ashraf M J

机构信息

Department of Immunology, Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.

出版信息

Transpl Infect Dis. 2015 Feb;17(1):21-4. doi: 10.1111/tid.12319. Epub 2014 Nov 29.

Abstract

OBJECTIVE

Umbilical cord blood (UCB) has been a reasonable alternative to granulocyte colony-stimulating factor-mobilized peripheral blood or bone marrow, as a source of hematopoietic stem cells with a lower risk of graft-versus-host disease. In immunocompromised hosts after transplantation, the risk of viral infection in adults, especially with beta-herpesviruses such as human herpesvirus-7 (HHV-7), may be increased. This virus in immunocompromised patients can be reactivated from latency and converted to an active phase. Therefore, light-upon-extension real-time polymerase chain reaction (PCR) was developed to assess the prevalence and load of HHV-7 in the plasma and buffy coat of donors.

METHODS

About 825 UCB samples under standard protocol from donors were collected. Then, DNA from plasma and buffy coat was extracted and quantitative real-time PCR was performed with light-upon-extension primers.

RESULTS

Overall, HHV-7 was detected in 3.64% (30/825) of UCB donors. HHV-7 DNA was detected in 26 (3.2%) buffy coat samples (latent infection), and only 4 (0.48%) of them were positive for HHV-7 DNA in plasma samples (active infection); the mean HHV-7 viral load was 1.31 × 10(1) copies/mL in latent infection, and 1.94 × 10(5) copies/mL in active infection.

CONCLUSIONS

We suggest that real-time PCR in plasma and buffy coat could be a useful method to detect active and latent HHV-7 infection in UCB donors and determine its role in subsequent transmission events.

摘要

目的

脐带血(UCB)已成为粒细胞集落刺激因子动员的外周血或骨髓的合理替代物,作为造血干细胞的来源,其移植物抗宿主病风险较低。在移植后的免疫功能低下宿主中,成人病毒感染的风险可能会增加,尤其是感染β疱疹病毒,如人类疱疹病毒7型(HHV-7)。免疫功能低下患者体内的这种病毒可从潜伏状态重新激活并转化为活跃期。因此,开发了延伸发光实时聚合酶链反应(PCR)来评估供体血浆和血沉棕黄层中HHV-7的流行率和载量。

方法

按照标准方案收集了约825份来自供体的脐带血样本。然后,提取血浆和血沉棕黄层中的DNA,并使用延伸发光引物进行定量实时PCR。

结果

总体而言,在825名脐带血供体中,有3.64%(30/825)检测到HHV-7。在26份(3.2%)血沉棕黄层样本中检测到HHV-7 DNA(潜伏感染),其中只有4份(0.48%)血浆样本中HHV-7 DNA呈阳性(活跃感染);潜伏感染时HHV-7病毒载量的平均值为1.31×10(1)拷贝/毫升,活跃感染时为1.94×10(5)拷贝/毫升。

结论

我们认为,血浆和血沉棕黄层中的实时PCR可能是检测脐带血供体中HHV-7活跃和潜伏感染并确定其在后续传播事件中作用的有用方法。

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