免疫功能低下的肺炎患者支气管肺泡灌洗液中巨细胞病毒的病毒培养和 DNA 定量检测。

Detection of cytomegalovirus in bronchoalveolar lavage fluid from immunocompromised patients with pneumonitis by viral culture and DNA quantification.

机构信息

Genetics and Microbiology Department. Universitat Autònoma de Barcelona, Spain; Microbiology Department, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain; Sant Pau Institute of Biomedical Research (IIb Sant Pau), Barcelona, Spain.

出版信息

J Virol Methods. 2023 Jul;317:114743. doi: 10.1016/j.jviromet.2023.114743. Epub 2023 Apr 26.

DOI:10.1016/j.jviromet.2023.114743

PMID:37116585

Abstract

PURPOSE

To compare the detection of human cytomegalovirus (HCMV) in bronchoalveolar lavage (BAL) fluid by viral culture and quantitative polymerase chain reaction (qPCR), and to establish a viral load threshold that can identify cases of HCMV replication indicative of pneumonitis. There is currently no universal viral load cut-off to differentiate between patients with and without pneumonitis, and the interpretation of qPCR results is challenging.

METHODS

176 consecutive BAL samples from immunosuppressed hosts with signs and/or symptoms of respiratory infection were prospectively studied by viral culture and qPCR.

RESULTS

Concordant results were obtained in 81.25% of the BAL samples. The rest were discordant, as only 34% of the qPCR-positive BAL samples were positive by culture. The median HCMV load was significantly higher in culture-positive than in culture-negative BAL samples (5038 vs 178 IU/mL). Using a cut-off value of 1258 IU/mL of HCMV in BAL, pneumonia was diagnosed with a sensitivity of 76%, a specificity of 100%, a VPP of 100% and VPN of 98%, and HCMV was isolated in 100% of the BAL cultures.

CONCLUSION

We found that a qPCR-negative was a quick and reliable way of ruling out HCMV pneumonitis, but a positive result did not always indicate clinically significant replication in the lung. However, an HCMV load in BAL fluid of ≥ 1258 IU/mL was always associated with disease, whereas < 200 IU/mL rarely so.

摘要

目的

比较病毒培养和实时聚合酶链反应（qPCR）检测支气管肺泡灌洗液（BAL）中人巨细胞病毒（HCMV）的效果，建立可识别提示肺炎的 HCMV 复制的病毒载量阈值。目前尚无用于区分有无肺炎的通用病毒载量临界值，且 qPCR 结果的解读具有挑战性。

方法

前瞻性研究了 176 例有呼吸道感染症状和/或体征的免疫抑制宿主的连续 BAL 样本，同时进行病毒培养和 qPCR。

结果

81.25%的 BAL 样本结果一致。其余样本结果不一致，因为 qPCR 阳性的 BAL 样本中只有 34%通过培养阳性。培养阳性的 BAL 样本中的 HCMV 载量中位数明显高于培养阴性的 BAL 样本（5038 与 178 IU/ml）。使用 BAL 中 HCMV 的 1258 IU/ml 截断值，肺炎的诊断敏感性为 76%，特异性为 100%，阳性预测值为 100%，阴性预测值为 98%，且 100%的 BAL 培养物可分离出 HCMV。