Newborn Blood Spot Screening for Sickle Cell Disease by Using Tandem Mass Spectrometry: Implementation of a Protocol to Identify Only the Disease States of Sickle Cell Disease
Newborn Blood Spot Screening for Sickle Cell Disease by Using Tandem Mass Spectrometry: Implementation of a Protocol to Identify Only the Disease States of Sickle Cell Disease
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DOI：
10.1373/clinchem.2013.210948
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发表时间：
2014-02-01
影响因子：
9.3
通讯作者：
Hillier, Sharon
中科院分区：
文献类型：
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作者：
Moat, Stuart J.;Rees, Derek;Hillier, Sharon
背景：目前推荐的新生儿镰状细胞病（SCD）血斑筛查技术采用高效液相色谱（HPLC）和等电聚焦技术，既能识别疾病状态又能识别携带者状态，导致大量婴儿受到不必要的随访。使用串联质谱 (MS/MS) 进行血斑胰蛋白酶肽分析是检测血红蛋白 (Hb) 变异性疾病的替代技术。 方法：我们在胰蛋白酶消化后使用 MS/MS 分析了 2154 个残留的新生儿血斑和 675 个 Hb 变异婴儿的新生儿血斑。通过使用 HbS、C、D-Punjab、O-Arab、Lepore 和 E 肽的变异肽与野生型肽丰度之间的比率来制定筛选截止值。使用这些截断值开发了分析后数据分析方案，仅检测 SCD 的疾病状态，而不识别携带者状态。通过 MS/MS 和 HPLC 对来自 SCD 高发地区的 13 249 个新生儿血点进行了平行研究。结果：制定的筛查临界值区分了患有 SCD 疾病状态的婴儿、SCD 携带者和 Hb 正常的婴儿。在平行研究中，没有发现假阴性结果，并且使用 MS/MS 方案正确识别了所有临床相关病例。揭盲数据显示，共有 328 名携带者婴儿被该方案成功排除。 结论：制定的筛查方案正确识别了患有 SCD 疾病状态的婴儿。此外，大量镰状细胞携带者婴儿未被成功识别，从而避免了不必要的后续检测和转诊遗传咨询。 (C) 2013年美国临床化学协会
BACKGROUND: The currently recommended technologies of HPLC and isoelectric focusing for newborn blood spot screening for sickle cell disease (SCD) identify both the disease and carrier states, resulting in large numbers of infants being followed up unnecessarily. Analysis of blood spot tryptic peptides performed by using tandem mass spectrometry (MS/MS) is an alternative technology to detect hemoglobin (Hb) variant disorders.METHODS: We analyzed 2154 residual newborn blood spots and 675 newborn blood spots from infants with Hb variants by using MS/MS after trypsin digestion. Screening cutoffs were developed by using the ratio between the variant peptide-to-wild-type peptide abundance for HbS, C, D-Punjab, O-Arab, Lepore, and E peptides. A postanalytical data analysis protocol was developed using these cutoffs to detect only the disease states of SCD and not to identify carrier states. A parallel study of 13 249 newborn blood spots from a high-prevalence SCD area were analyzed by both MS/MS and HPLC.RESULTS: Screening cutoffs developed distinguished the infants with the disease states of SCD, infants who were carriers of SCD, and infants with normal Hb. In the parallel study no false-negative results were identified, and all clinically relevant cases were correctly identified using the MS/MS protocol. Unblinding the data revealed a total of 328 carrier infants that were successfully excluded by the protocol.CONCLUSIONS: The screening protocol developed correctly identified infants with the disease states of SCD. Furthermore, large numbers of sickle cell carrier infants were successfully not identified, thereby avoiding unnecessary follow-up testing and referral for genetic counseling. (C) 2013 American Association for Clinical Chemistry