Cytomegalovirus DNA detection in Guthrie cards:: a powerful tool for diagnosing congenital infection
Cytomegalovirus DNA detection in Guthrie cards:: a powerful tool for diagnosing congenital infection
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DOI：
10.1016/s1386-6532(00)00089-5
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发表时间：
2000-09-01
影响因子：
8.8
通讯作者：
Melotti, D
中科院分区：
文献类型：
--
作者：
Barbi, M;Binda, S;Melotti, D
背景：先天性巨细胞病毒感染的简单可靠的诊断对于临床和流行病学都是必要的。这可以通过在古特里卡上的血斑（DBS）中显示巨细胞病毒（CMV）DNA来完成。目的：（1）评估与病毒分离相比，该方法（DBS试验）诊断先天性CMV感染的敏感性和特异性;（2）评估该试验在先天性CMV晚期诊断中的应用。研究设计：该方法在（1）509名婴儿（72例阳性）和（2）191名3周后（25天至5岁）的儿童的卡片上进行了测试。将血液从古特里卡片上洗脱并热提取。PCR扩增CMV糖蛋白B（gB）基因的一个区域，琼脂糖凝胶电泳检测扩增产物。结果如下：DBS试验在所有72例先天性感染的婴儿中均为阳性，在437例巨细胞病毒分离阴性的婴儿中有4例为阳性（敏感性100%，特异性99%）。在92名晚期病毒分离的婴儿中，有16名婴儿的感染被证明是先天性的，该测试还在83名未进行病毒培养的儿童中发现了18名先天性感染（13名有症状，5名无症状）。另外五十六例对照病例检测结果为阴性。结论：DBS试验是诊断先天性巨细胞病毒感染的可靠方法，可作为病毒培养的替代方法。它能够揭示确定的CMV感染是先天性的还是在病毒分离无法做到这一点的年龄出生后。它可以评估输血等危险程序的作用，并可以确定诊断较晚或起源有争议的疾病的病因。(C)出版社：Elsevier Science B. V.
Background: A simple and reliable diagnosis of congenital cytomegalovirus infection is necessary both for clinical and epidemiological purposes. This could be accomplished through the demonstration of cytomegalovirus (CMV) DNA in blood spots (DBS) on Guthrie cards. Objectives: (I) To assess the sensitivity and specificity of the method (DBS test) in diagnosing congenital CMV infection compared with viral isolation and (2) to evaluate the applications of the test to the late diagnosis of congenital CMV. Study design: The method was tested on the cards of (1) 509 babies examined through viral isolation within their third week of life (72 positive cases) and (2) 191 children studied after 3 weeks of life (25 days to 5 years). Blood was eluted from Guthrie cards and heat extracted. The products of a nested polymerase chain reaction (PCR) amplifying one region in the CMV glycoprotein B (gB) gene were detected by agarose gel electrophoresis. Results: DBS test was positive in all 72 congenitally infected babies and in four of the 437 negative at cytomegalovirus isolation (sensitivity 100%, specificity 99%). Infection in 16 of the 92 infants with a late viral isolation was demonstrated to be congenital by the test, which also detected congenital infection in 18 of 83 children in whom viral culture was not performed (13 with and five without symptoms). Fifty-six additional control cases tested negative. Conclusions: DBS test is a reliable assay for diagnosing congenital cytomegalovirus infection and could be used as an alternative to viral culture. It is able to reveal whether ascertained CMV infection is congenital or postnatal at an age when viral isolation is not able to do so. It can assess the role of risky procedures such as transfusion and it can ascertain the etiology of morbid conditions diagnosed late or of controversial origin. (C) 2000 Published by Elsevier Science B.V.