A Novel Diagnostic Tool for Detecting Neonatal Infections Using Multiplex Polymerase Chain Reaction
A Novel Diagnostic Tool for Detecting Neonatal Infections Using Multiplex Polymerase Chain Reaction
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DOI：
10.1159/000208791
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发表时间：
2009-01-01
期刊：
影响因子：
2.5
通讯作者：
Matsuo, Masafumi
中科院分区：
文献类型：
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作者：
Enomoto, Masahiro;Morioka, Ichiro;Matsuo, Masafumi
背景资料：在新生儿感染中，有必要快速准确地检测各种病原体，因为当诊断延误时，感染往往是致命的。然而，没有诊断工具，快速检测病原体引起的新生儿传染病。目的：目的建立一种多重聚合酶链反应（PCR）快速诊断新生儿感染的8种主要病原体（B族链球菌、大肠埃希菌、铜绿假单胞菌、耐甲氧西林金黄色葡萄球菌、解脲脲原体、单纯疱疹病毒、巨细胞病毒和白色念珠菌）的方法，并在新生儿重症监护病房（NICU）进行验证。方法：130例临床样本来自有任何感染体征或病史的新生儿。从这些样品中提取DNA，并用8种引物对的混合物进行多重PCR，所有引物对均设计用于扩增致病性DNA并产生不同大小的扩增子。同时对77份疑似细菌感染的标本进行细菌培养，以评价多重PCR检测结果的准确性。结果：多重PCR方法可在3.5- 4.5h内快速检测出8种病原菌中的6种。这些积极的结果使我们能够立即诊断并选择针对每种病原体的适当药物。与培养结果比较，我们的检测特征如下：特异性：93%，阴性预测值：96%，一致率：90%。结论：我们已经建立并验证了一种快速诊断工具，用于检测病原体，使用多重PCR，这可能是有用的新生儿感染的确诊NICU。版权所有（C）2009 S. Karger AG，巴塞尔
Background: In newborns with infections, it is necessary to detect various pathogens rapidly and accurately, because the infections are often fatal when diagnosis is delayed. However, no diagnostic tools that rapidly detect pathogens causing neonatal infectious diseases are available. Objectives: To establish a rapid diagnostic tool using multiplex polymerase chain reaction (PCR) to detect 8 major pathogens that often cause neonatal infections, including Group B Streptococcus, Escherichia coli, Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, Ureaplasma urealyticum, herpes simplex virus, Cytomegalovirus, and Candida albicans, and to validate this tool in the neonatal intensive care unit (NICU). Methods: One hundred and thirty clinical samples were obtained from newborns with any infectious signs or histories. DNA was extracted from these samples and multiplex PCR was performed with a mixture of 8 primer pairs, all designed to amplify pathogenic DNA and produce different sizes of amplicons. Seventy-seven samples with suspicion of bacterial infections were also examined by bacterial culture to evaluate the accuracy of the multiplex PCR results. Results: Six of the 8 pathogens could be rapidly detected by our multiplex PCR method, within 3.5-4.5 h. These positive results led us to immediately diagnose and select proper drugs against each pathogen. In comparison with culture results, our test characteristics were as follows: specificity: 93%, negative predictive value: 96%, and concordance rate: 90%. Conclusions: We have established and validated a rapid diagnostic tool for detecting pathogens using multiplex PCR, which may be useful for the confirmed diagnosis of neonatal infections in the NICU. Copyright (C) 2009 S. Karger AG, Basel