Characterization of a universal screening approach for congenital CMV infection based on a highly-sensitive, quantitative, multiplex real-time PCR assay
Characterization of a universal screening approach for congenital CMV infection based on a highly-sensitive, quantitative, multiplex real-time PCR assay
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DOI：
10.1371/journal.pone.0227143
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发表时间：
2020-01-09
期刊：
影响因子：
3.7
通讯作者：
Ueberla, Klaus
中科院分区：
文献类型：
--
作者：
Nagel, Angela;Dimitrakopoulou, Emmanouela;Ueberla, Klaus
大多数先天性巨细胞病毒（cCMV）感染在出生时无症状，因此无法诊断。这些婴儿中约有10-15%会出现迟发性听力损失和其他发育障碍。在出生时实施普遍筛查方法，可通过对高危婴儿进行更密切的随访，及早开始对症干预，并提供机会考虑治疗晚发性疾病。与快速培养筛选法相比，用于检测口腔拭子样本中CMV DNA的实时PCR法证明了可行性和良好的临床灵敏度。由于大多数cCMV感染保持无症状，根据迟发性cCMV疾病的低风险和高风险对CMV感染婴儿进行分层的通用筛查测定可以限制父母的焦虑并降低随访成本。因此，我们开发并表征了一种基于高灵敏度定量实时PCR测定的筛选算法，其与来自通用筛选的样品的集中测试兼容，并且允许以国际单位（IU）/ml唾液或IU/10（5）细胞当量来确定唾液样品的CMV DNA载量。通过筛查算法检测呈阳性的34份新生儿唾液样本中有18份通过检测生命最初几周内获得的血液和/或尿液样本中的CMV DNA得到确认。所有无法确认的筛查样本的病毒载量< 2.3x10（5）IU/ml唾液（中位数：6.8x10（3））或1.3x10（5）IU/10（5）细胞当量（中位数：4.0x10（2））。确认cCMV感染的筛选样本的病毒载量范围为7.5x10（2）至8.2x10（9）IU/ml唾液（中位数：9.3x10（7））或1.5x10（2）至5.6x10（10）IU/10（5）细胞当量（中位数：3.5x10（6））。对这些确诊为cCMV感染的新生儿进行临床随访，应揭示晚发型cCMV疾病的风险是否与早期唾液样本中的CMV DNA载量相关，以及是否可以定义一个临界值，以识别有或无晚发型cCMV疾病风险的cCMV感染婴儿。
The majority of congenital cytomegalovirus (cCMV) infections are asymptomatic at birth and therefore not diagnosed. Approximately 10-15% of these infants develop late-onset hearing loss and other developmental disorders. Implementation of a universal screening approach at birth may allow early initiation of symptomatic interventions due to a closer follow-up of infants at risk and offers the opportunity to consider treatment of late-onset disease. Realtime PCR assays for the detection of CMV DNA in buccal swab samples demonstrated feasibility and good clinical sensitivity in comparison to a rapid culture screening assay. Because most cCMV infections remain asymptomatic, a universal screening assay that stratifies CMV infected infants according to low and high risk of late-onset cCMV disease could limit the parental anxiety and reduce follow-up costs. We therefore developed and characterized a screening algorithm based on a highly-sensitive quantitative real-time PCR assay that is compatible with centralized testing of samples from universal screening and allows to determine CMV DNA load of saliva samples either as International Units (IU)/ml saliva or IU/10(5) cell equivalents. 18 of 34 saliva samples of newborns that tested positively by the screening algorithm were confirmed by detection of CMV DNA in blood and/or urine samples obtained during the first weeks of life. All screening samples that could not be confirmed had viral loads of < 2.3x10(5) IU/ml saliva (median: 6.8x10(3)) or 1.3x10(5) IU/10(5) cell equivalents (median: 4.0x10(2)). The viral load of screening samples with confirmed cCMV infection ranged from 7.5x10(2) to 8.2x10(9) IU/ml saliva (median: 9.3x10(7)) or 1.5x10(2) to 5.6x10(10) IU/10(5) cell equivalents (median: 3.5x10(6)). Clinical follow-up of these newborns with confirmed cCMV infection should reveal whether the risk of late-onset cCMV disease correlates with CMV DNA load in early life saliva samples and whether a cut-off can be defined identifying cCMV infected infants with or without risk for late-onset cCMV disease.