Detection and quantification of human metapneumovirus in pediatric specimens by real-time RT-PCR.
Detection and quantification of human metapneumovirus in pediatric specimens by real-time RT-PCR.
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DOI：
10.1016/j.jcv.2004.11.023
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发表时间：
2005-08
期刊：
影响因子：
--
通讯作者：
Morrow R
中科院分区：
文献类型：
--
作者：
Kuypers J;Wright N;Corey L;Morrow R
人类偏肺病毒(HMPV)是一种新近发现的病毒，可导致儿童呼吸道疾病。建立了实时逆转录聚合酶链式反应(RT-PCR)检测呼吸道标本中hMPV的方法。定量RT-PCR从hMPV融合蛋白基因中扩增出约70个碱基对的片段。该化验方法经过验证，并用于测试2002年12月至2003年5月期间在华盛顿州西雅图一家医院就诊的儿童的呼吸道样本。该方法检测到1000个hMPV拷贝/毫升样本，没有检测到19种其他呼吸道病毒，并能够在由20个先前测试的样本组成的熟练小组中检测和准确量化来自四个已知hMPV遗传谱系的分离物。在719例呼吸道标本中，52例(7.2%)检出hMPV。52份阳性标本中hMPV 10拷贝数/毫升的平均对数为7.67(范围：0.49~10.60)。7-12个月儿童的HMPV感染率(12.4%)显著高于7个月以下儿童(4.7%)(P<0.005)。该年龄组儿童呼吸道标本中hMPV水平(平均对数为8.43拷贝/毫升)也明显高于年龄较小的儿童(平均对数为6.93拷贝/毫升)(P=0.0025)。本文所述的快速实时RT-PCR检测方法是一种敏感的检测方法，可用于阐明hMPV的流行病学和与其相关的疾病。
Human metapneumovirus (hMPV), a recently identified virus, causes respiratory illness in children. A real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was developed and used to detect and quantify hMPV in respiratory specimens. The quantitative RT-PCR assay amplified an approximately 70 base pair fragment from the hMPV fusion protein gene. The assay was validated and used to test respiratory specimens obtained from children seen at a hospital in Seattle, Washington, from December 2002 through May 2003. The assay detected 1000 hMPV copies/mL of specimen, did not detect 19 other respiratory viruses, and was able to detect and accurately quantify isolates from the four known hMPV genetic lineages in a proficiency panel of 20 previously tested samples. hMPV was detected in 52 (7.2%) of 719 pediatric respiratory specimens. The mean log 10 copies/mL of hMPV in the 52 positive specimens was 7.67 (range = 4.59–10.60). Children aged 7–12 months had a significantly higher hMPV prevalence (12.4%) than did children younger than 7 months (4.7%) (P < 0.005). Children in this age group also had significantly higher levels of hMPV in their respiratory specimens (mean log 8.43 copies/mL) than did the younger children (mean log 6.93 copies/mL) (P = 0.0025). The rapid real-time RT-PCR assay described here is a sensitive test for clarifying the epidemiology of and diseases associated with hMPV.
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影响因子：
82.9
作者：
van den Hoogen BG;de Jong JC;Groen J;Kuiken T;de Groot R;Fouchier RA;Osterhaus AD
通讯作者：
Osterhaus AD
影响因子：
11.8
作者：
Mullins JA;Erdman DD;Weinberg GA;Edwards K;Hall CB;Walker FJ;Iwane M;Anderson LJ
通讯作者：
Anderson LJ
DOI：
10.1016/j.jcv.2004.03.018
发表时间：
2004-10
期刊：
Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
影响因子：
--
作者：
Kuypers J;Wright N;Morrow R
通讯作者：
Morrow R
影响因子：
9.4
作者：
Maertzdorf, J;Wang, CK;Fouchier, RAM
通讯作者：
Fouchier, RAM
影响因子：
6.4
作者：
Falsey, AR;Erdman, D;Walsh, EE
通讯作者：
Walsh, EE