Dried Blood Spot Real-time Polymerase Chain Reaction Assays to Screen Newborns for Congenital Cytomegalovirus Infection
Dried Blood Spot Real-time Polymerase Chain Reaction Assays to Screen Newborns for Congenital Cytomegalovirus Infection
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DOI：
10.1001/jama.2010.423
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发表时间：
2010-04-14
影响因子：
120.7
通讯作者：
Fowler, Karen B.
中科院分区：
文献类型：
--
作者：
Boppana, Suresh B.;Ross, Shannon A.;Fowler, Karen B.
背景：需要可靠的方法筛查新生儿先天性巨细胞病毒（CMV）感染，以识别听力损失风险增加的婴儿。由于干血斑（DBS）是常规收集的代谢筛查从所有新生儿在美国，有兴趣在使用DBS聚合酶链反应（PCR）为基础的方法为新生儿CMVscreen.Objective新生儿CMVscreen.Design，设置，和参与者在2007年3月和2008年5月之间，在7个美国医疗中心出生的婴儿通过快速培养测试唾液样本的早期抗原荧光灶。将唾液快速培养的结果与单引物（2007年3月至2007年12月）和双引物DBS实时PCR（2008年1月至2008年5月）进行比较。样本经快速培养或PCR筛选为阳性的婴儿，通过唾液或尿液快速培养检测的参考标准方法确认为先天性感染。单引物和双引物DBS真实的-结果20448例患儿中，92例确诊为先天性CMV感染，其中10例患儿中，12例为先天性CMV感染（0.45%; 95%置信区间[CI]，0.36%-0.55%）婴儿。92名婴儿中有91名唾液快速培养结果阳性。在使用单引物DBS PCR筛查的11422名婴儿中，60名婴儿中有17名（28%）婴儿的检测结果呈阳性，而在使用双引物DBS PCR筛查的9026名婴儿中，32名婴儿中有11名（34%）筛查结果呈阳性。单引物DBS PCR鉴定先天性CMV感染的灵敏度为28.3%（95%CI，17.4%-41.4%），特异性为99.9%（95%CI，99.9%-100%），阳性LR为803.7（95%CI，278.7-2317.9），阴性LR为0.7（95%CI，0.60.8）。单引物DBS PCR的阳性和阴性预测值分别为80.9%（95%CI，58.1%-94.5%）和99.6%（95%CI，99.5%-99.7%）。双引物DBS PCR检测先天性CMV感染的敏感性为34.4%（95% CI，18.6%-53.2%），特异性为99.9%（95% CI，99.9%-100.0%），阳性LR为3088.9（95% CI，410.8-23 226.7），阴性LR为0.7（95% CI，0.5-0.8）。结论DBS实时荧光定量PCR检测新生儿巨细胞病毒敏感性较唾液快速培养低，限制了其作为新生儿巨细胞病毒筛查的价值。美国医学会杂志2010; 303（14）：1375-1382
Context Reliable methods to screen newborns for congenital cytomegalovirus (CMV) infection are needed for identification of infants at increased risk of hearing loss. Since dried blood spots (DBS) are routinely collected for metabolic screening from all newborns in the United States, there has been interest in using DBS polymerase chain reaction (PCR)-based methods for newborn CMV screening.Objective To determine the diagnostic accuracy of DBS real-time PCR assays for newborn CMV screening.Design, Setting, and Participants Between March 2007 and May 2008, infants born at 7 US medical centers had saliva specimens tested by rapid culture for early antigen fluorescent foci. Results of saliva rapid culture were compared with a single-primer (March 2007-December 2007) and a 2-primer DBS real-time PCR (January 2008-May 2008). Infants whose specimens screened positive on rapid culture or PCR had congenital infection confirmed by the reference standard method with rapid culture testing on saliva or urine.Main Outcome Measures Sensitivity, specificity, and positive and negative likelihood ratios (LRs) of single-primer and 2-primer DBS real-time PCR assays for identifying infants with confirmed congenital CMV infection.Results Congenital CMV infection was confirmed in 92 of 20 448 (0.45%; 95% confidence interval [CI], 0.36%-0.55%) infants. Ninety-one of 92 infants had positive results on saliva rapid culture. Of the 11 422 infants screened using the single-primer DBS PCR, 17 of 60 (28%) infants had positive results with this assay, whereas, among the 9026 infants screened using the 2-primer DBS PCR, 11 of 32 (34%) screened positive. The single-primer DBS PCR identified congenital CMV infection with a sensitivity of 28.3% (95% CI, 17.4%-41.4%), specificity of 99.9% (95% CI, 99.9%-100%), positive LR of 803.7 (95% CI, 278.7-2317.9), and negative LR of 0.7 (95% CI, 0.60.8). The positive and negative predictive values of the single-primer DBS PCR were 80.9% (95% CI, 58.1%-94.5%) and 99.6% (95% CI, 99.5%-99.7%), respectively. The 2-primer DBS PCR assay identified infants with congenital CMV infection with a sensitivity of 34.4% (95% CI, 18.6%-53.2%), specificity of 99.9% (95% CI, 99.9%100.0%), positive LR of 3088.9 (95% CI, 410.8-23 226.7), and negative LR of 0.7 (95% CI, 0.5-0.8). The positive and negative predictive values of the 2-primer DBS PCR were 91.7% (95% CI, 61.5%-99.8%) and 99.8% (95% CI, 99.6%-99.9%), respectively.Conclusion Among newborns, CMV testing with DBS real-time PCR compared with saliva rapid culture had low sensitivity, limiting its value as a screening test. JAMA. 2010; 303(14): 1375-1382