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比利时健康献血者中人类细小病毒B19 DNA的流行率及其与针对病毒结构和非结构蛋白的特异性抗体的相关性

Prevalence of human erythrovirus B19 DNA in healthy Belgian blood donors and correlation with specific antibodies against structural and non-structural viral proteins.

作者信息

Thomas I, Di Giambattista M, Gérard C, Mathys E, Hougardy V, Latour B, Branckaert T, Laub R

机构信息

Department of Microbiology, Scientific Institute of Public Health, Brussels, Belgium.

出版信息

Vox Sang. 2003 May;84(4):300-7. doi: 10.1046/j.1423-0410.2003.00299.x.

Abstract

BACKGROUND AND OBJECTIVES

Human parvovirus (erythrovirus) B19 is recognized as a major contaminant of blood and blood products. To reduce the risk of contamination, plasma-pool screening and exclusion of highly viraemic donations are recommended. The objectives of this study were to estimate the prevalence of B19 DNA in our blood-donor population, to determine the appropriate pool size to be tested (taking into account parameters such as prevalence, viral load, test sensitivity, and the efficacy of inactivation procedures), and to correlate viral loads with the serological status of donors as regards antibodies against different viral proteins.

MATERIALS AND METHODS

Pools of different sizes were tested for B19, using a sensitive nested polymerase chain reaction (PCR) as well as an simple, un-nested, less sensitive PCR. Positive pools were resolved to the level of individual donations, and the viral load and serological markers were determined.

RESULTS

Of 16,859 donations, 27 (one of 625) were found to be B19 DNA positive, with viral loads ranging from 10(2) to > 10(7) IU/ml. Twenty-five of the positive donations were tested for VP-specific anti-B19 antibodies, and eight (32%) were negative for both immunoglobulin (Ig)M and IgG. They were probably collected in the preseroconversion window period or from chronic carriers without detectable antibodies. We regarded the seven (28%) IgM-positive donors as being in the early phase of infection. The remaining 10 (40%) IgM-negative, IgG-positive donors were probably carriers of persistent infection (i.e. PCR positive despite the presence of IgG antibodies), as suggested by their low viral loads (< 10(4) IU/ml). Fifteen out of 36 major pools contained one or more contaminated donations. Among these, 12 tested positive by nested PCR and only three by un-nested PCR, this reflecting a viral load of > 10(4) IU/ml.

CONCLUSIONS

By testing all donations as pools of 480 by un-nested PCR, and resolving positive pools to identify the responsible donations, it is possible to ensure that the viral load in fractionation pools (5000 donations) remains < 10(3) IU/ml, compatible with the efficacy of inactivation procedures and complying with Food and Drug Administration (FDA) recommendations.

摘要

背景与目的

人类细小病毒(红病毒)B19被认为是血液及血液制品的主要污染物。为降低污染风险,建议进行血浆池筛查并排除病毒血症水平高的献血。本研究的目的是估计我们献血人群中B19 DNA的流行率,确定合适的待检测池大小(考虑流行率、病毒载量、检测灵敏度及灭活程序的效力等参数),并将病毒载量与献血者针对不同病毒蛋白的抗体血清学状态相关联。

材料与方法

使用灵敏的巢式聚合酶链反应(PCR)以及简单的、非巢式的、灵敏度较低的PCR对不同大小的池进行B19检测。阳性池被分解至个体献血水平,并测定病毒载量和血清学标志物。

结果

在16859份献血中,27份(625份中的1份)被发现B19 DNA呈阳性,病毒载量范围为10²至>10⁷ IU/ml。对25份阳性献血检测了VP特异性抗B19抗体,其中8份(32%)的免疫球蛋白(Ig)M和IgG均为阴性。它们可能是在血清转化前窗口期采集的,或者来自无可检测抗体的慢性携带者。我们将7份(28%)IgM阳性的献血者视为处于感染早期。其余10份(40%)IgM阴性、IgG阳性的献血者可能是持续性感染的携带者(即尽管存在IgG抗体但PCR仍呈阳性),这从其低病毒载量(<10⁴ IU/ml)可以看出。36个主要血浆池中,有15个含有一份或多份受污染的献血。其中,12个通过巢式PCR检测呈阳性,只有3个通过非巢式PCR检测呈阳性,这反映病毒载量>10⁴ IU/ml。

结论

通过用非巢式PCR将所有献血作为480份的池进行检测,并分解阳性池以识别责任献血,可以确保分馏池(5000份献血)中的病毒载量保持<10³ IU/ml,这与灭活程序的效力相符且符合美国食品药品监督管理局(FDA)的建议。

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