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通过DNA扩增早期检测骨髓移植受者中的人巨细胞病毒血症

Early detection of human cytomegalovirus viremia in bone marrow transplant recipients by DNA amplification.

作者信息

Nolte F S, Emmens R K, Thurmond C, Mitchell P S, Pascuzzi C, Devine S M, Saral R, Wingard J R

机构信息

Department of Pathology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.

出版信息

J Clin Microbiol. 1995 May;33(5):1263-6. doi: 10.1128/jcm.33.5.1263-1266.1995.

Abstract

Surveillance blood cultures for human cytomegalovirus (HCMV) are commonly used to identify the bone marrow transplant (BMT) recipients with the highest risk of serious HCMV disease and for whom early interventional ganciclovir therapy would be beneficial. We monitored 36 allogeneic BMT recipients weekly for the presence of HCMV in the blood from 0 to 100 days posttransplantation. Viable HCMV in leukocytes (WBC) was detected by shell vial and tube culture methods. HCMV DNA in WBC and plasma was detected by PCR and DNA hybridization using primers and a probe from the EcoRI fragment D region of HCMV AD169. A uracil-N-glycosylase-dUTP PCR protocol was used to prevent false-positive results due to amplicon carryover. Seventeen patients had multiple consecutive positive samples containing HCMV DNA in plasma or WBC. In 14 of 17 patients, HCMV was also detected by blood culture. HCMV DNA was detected sporadically in six patients, none of whom had positive cultures. One patient had HCMV viremia detected by WBC culture only. The remaining 12 patients had no positive PCR assays or blood cultures. For the patients with positive blood cultures, PCR detection of HCMV DNA in plasma preceded detection of HCMV in culture by a mean of 8 days and detection in WBC preceded detection in culture by 6 days. HCMV disease (interstitial pneumonia) was documented for two patients with viremia (blood culture and PCR positive) and one patient without viremia (blood culture and PCR negative). The earlier recognition of high-risk patients provided by detection of HCMV DNA in plasma or WBC may improve the efficacy of early interventional antiviral therapy.

摘要

对人类巨细胞病毒(HCMV)进行监测性血培养常用于识别骨髓移植(BMT)受者中发生严重HCMV疾病风险最高的患者,以及早期进行更昔洛韦干预性治疗可能有益的患者。我们对36例异基因BMT受者在移植后0至100天每周监测一次血液中是否存在HCMV。采用空斑小室法和试管培养法检测白细胞(WBC)中的活HCMV。使用来自HCMV AD169的EcoRI片段D区域的引物和探针,通过PCR和DNA杂交检测WBC和血浆中的HCMV DNA。采用尿嘧啶-N-糖基化酶-dUTP PCR方案以防止由于扩增子残留导致的假阳性结果。17例患者有多个连续的血浆或WBC样本呈HCMV DNA阳性。17例患者中有14例血培养也检测到HCMV。6例患者偶尔检测到HCMV DNA,他们均无培养阳性结果。1例患者仅通过WBC培养检测到HCMV病毒血症。其余12例患者PCR检测和血培养均为阴性。对于血培养阳性的患者,血浆中HCMV DNA的PCR检测比培养检测到HCMV平均提前8天,WBC中检测到HCMV比培养检测提前6天。记录到2例病毒血症患者(血培养和PCR阳性)和1例无病毒血症患者(血培养和PCR阴性)发生了HCMV疾病(间质性肺炎)。通过检测血浆或WBC中的HCMV DNA更早识别高危患者可能会提高早期干预性抗病毒治疗的疗效。

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