肺移植受者血浆和外周血淋巴细胞中巨细胞病毒（HCMV）DNA的检测与定量：COBAS Amplicor HCMV监测检测法与内部定量HCMV聚合酶链反应的比较

HCMV DNA detection and quantitation in the plasma and PBL of lung transplant recipients: COBAS Amplicor HCMV monitor test versus in-house quantitative HCMV PCR.

作者信息

Michaelides Alexandra, Facey David, Spelman Denis, Wesselingh Steven, Kotsimbos Thomas

机构信息

Department of Microbiology and Infectious Diseases Unit, Alfred Hospital, Commercial Road, Prahran 3181, VIC, Australia.

出版信息

J Clin Virol. 2003 Oct;28(2):111-20. doi: 10.1016/s1386-6532(02)00272-x.

DOI:10.1016/s1386-6532(02)00272-x

PMID:12957181

Abstract

BACKGROUND

Human cytomegalovirus (HCMV) reactivation may cause severe disease in immunosuppressed patients. Quantitation of HCMV viral load in the blood has been shown to be important in predicting for HCMV disease, however the particular blood compartment that should be tested, plasma versus peripheral blood leukocytes (PBL), has been subject to debate.

OBJECTIVES

To simultaneously compare HCMV viral loads in the PBL using an in-house quantitative HCMV polymerase chain reaction (PCR) assay and in the plasma using the commercially available COBAS Amplicor HCMV monitor test, in a cohort of lung transplant recipients (LTR).

STUDY DESIGN

Sequential paired plasma and PBL samples were collected (n=98) from a total of 21 LTR during the first 6 months post lung transplantation. HCMV viral loads were assessed in the PBL using an in-house quantitative HCMV PCR assay and the plasma using the COBAS Amplicor HCMV monitor test. HCMV disease in LTR was defined as histopathologically proven HCMV pneumonitis.

RESULTS

HCMV deoxyribonucleic acid (DNA) was detected in 39/98 (40%) of total samples with excellent agreement between the two strategies. HCMV was detected in both the plasma and PBL in 38/39 (97%) of HCMV positive samples. HCMV viral loads were higher in patients with HCMV pneumonitis in both the PBL and plasma compared to patients without HCMV pneumonitis. Greater than 10-fold increases in HCMV DNA levels had a sensitivity of 88% and a specificity of 73% in the plasma for HCMV pneumonitis and a positive and negative predictive value of 70 and 89%, respectively. Greater that 10-fold increases in HCMV DNA levels in the PBL had a sensitivity of 88% and a specificity of 64%, for HCMV pneumonitis and a positive and negative predictive value of 64 and 88%, respectively.

CONCLUSIONS

Acknowledging the difference in assay methods used for HCMV DNA quantitation, the in-house PCR PBL assay and the COBAS Amplicor HCMV monitor plasma assay predicted equally well for HCMV pneumonitis in LTR.

摘要

背景

人巨细胞病毒（HCMV）再激活可能在免疫抑制患者中引发严重疾病。血液中HCMV病毒载量的定量已被证明在预测HCMV疾病方面很重要，然而，应该检测的特定血液成分，即血浆还是外周血白细胞（PBL），一直存在争议。

目的

在一组肺移植受者（LTR）中，使用内部定量HCMV聚合酶链反应（PCR）检测法同时比较PBL中的HCMV病毒载量，并使用市售的COBAS Amplicor HCMV监测检测法比较血浆中的HCMV病毒载量。

研究设计

在肺移植后的前6个月内，从总共21名LTR中顺序收集配对的血浆和PBL样本（n = 98）。使用内部定量HCMV PCR检测法评估PBL中的HCMV病毒载量，使用COBAS Amplicor HCMV监测检测法评估血浆中的HCMV病毒载量。LTR中的HCMV疾病定义为经组织病理学证实天天美剧网的HCMV肺炎。

结果

在39/98（40%）的总样本中检测到HCMV脱氧核糖核酸（DNA），两种检测策略之间具有良好的一致性。在38/39（97%）的HCMV阳性样本的血浆和PBL中均检测到HCMV。与没有HCMV肺炎的患者相比，患有HCMV肺炎的患者的PBL和血浆中的HCMV病毒载量更高。血浆中HCMV DNA水平升高超过10倍对HCMV肺炎的敏感性为88%，特异性为73%，阳性预测值和阴性预测值分别为70%和89%。PBL中HCMV DNA水平升高超过10倍对HCMV肺炎的敏感性为88%，特异性为64%，阳性预测值和阴性预测值分别为64%和88%。