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[荧光定量聚合酶链反应技术检测人巨细胞病毒感染及其在人巨细胞病毒感染儿童诊断与治疗中的应用]

[Detection of human cytomegalovirus infection by FQ-PCR technique and its application in the diagnosis and treatment of HCMV infected children].

作者信息

Chen Fenhua, He Zhengxian, Pan Sinian, Ning Fangqin, Wang Qingwen, Xiao Zuoyuan

机构信息

Department of Pediatrics, The Third Affiliated Hospital, Sun Yat Sen University, Guangzhou 510630, China.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2002 Jun;16(2):187-90.

Abstract

BACKGROUND

To detect quantitatively HCMV DNA in peripheral blood leukocytes to monitor the status of HCMV infection, evaluate the effectiveness of antiviral treatment with ganciclovir (GCV) combined with intravenous immunoglobulin (IVIG) and find out the relationship among the HCMV DNA levels, the state of infection and the clinical outcome.The long-term goal of the study was to establish a molecular diagnostic standard for HCMV infection in children.

METHODS

45 cases of suspected HCMV-infected children were examined by PCR, ELISA and fluorescent quantitative (FQ)-PCR, respectively. Twenty five HCMV hepatitis cases of the 45 were randomly assigned to a treated group or a control group. Both groups were treated with prednisone, glucurone, Luminal and Xiaoyanlidanpian. But the treated group was given with GCV+IVIG in addition. Each infant of the two groups was checked with FQ-PCR at the five time points.

RESULTS

The positive rates of PCR, ELISA and FQ-PCR were 60.00%, 33.33% and 66.67%,their sensitivities were 84.38%, 46.88% and 93.75%, respectively. There was no significant difference in viral DNA copy numbers between the two groups before being treated (P>0.05), but there was significant difference between HCMV hepatitis and normal infants (P<0.001). Although viral load of both groups decreased in both groups, the viral load of the treated group decreased more significantly. The level of HCMV DNA fell to 103 copies/ml at second time point while that of the control group fell to the same level after third time point. The differences between the two groups at each time point were statistically significant (P<0.001). The results of 135 person times testing indicated that 103 copies/ml of FQ-PCR can be taken as a critical value for prediction of active HCMV infection.

CONCLUSIONS

FQ-PCR may be one of the effective methods for diagnosis of HCMV disease; it can offer a key index in the diagnosis of HCMV active infection; dynamic detection of HCMV viral load can play a role not only in monitoring antiviral therapy, but also in evaluating the development and prognosis of HCMV disease.

摘要

背景

定量检测外周血白细胞中人类巨细胞病毒(HCMV)DNA,以监测HCMV感染状况,评估更昔洛韦(GCV)联合静脉注射免疫球蛋白(IVIG)抗病毒治疗的效果,并明确HCMV DNA水平、感染状态与临床结局之间的关系。本研究的长期目标是建立儿童HCMV感染的分子诊断标准。

方法

分别采用聚合酶链反应(PCR)、酶联免疫吸附测定(ELISA)和荧光定量(FQ)-PCR对45例疑似HCMV感染儿童进行检测。45例HCMV肝炎患儿中,25例被随机分为治疗组和对照组。两组均给予泼尼松、葡醛内酯、鲁米那和消炎利胆片治疗。但治疗组额外给予GCV+IVIG。两组患儿在5个时间点均进行FQ-PCR检测。

结果

PCR、ELISA和FQ-PCR的阳性率分别为60.00%、33.33%和66.67%,其敏感性分别为84.38%、46.88%和93.75%。两组治疗前病毒DNA拷贝数无显著差异(P>0.05),但HCMV肝炎患儿与正常婴儿之间存在显著差异(P<0.001)。虽然两组病毒载量均下降,但治疗组病毒载量下降更显著。治疗组在第二个时间点HCMV DNA水平降至10³拷贝/ml,而对照组在第三个时间点降至相同水平。两组在各时间点的差异具有统计学意义(P<0.001)。135人次检测结果表明,FQ-PCR检测10³拷贝/ml可作为预测HCMV活动性感染的临界值。

结论

FQ-PCR可能是诊断HCMV疾病的有效方法之一;它可为HCMV活动性感染的诊断提供关键指标;动态检测HCMV病毒载量不仅可用于监测抗病毒治疗,还可用于评估HCMV疾病的发展和预后。

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