Comparison of real-time PCR assays with fluorescent-antibody assays for diagnosis of respiratory virus infections in children
Comparison of real-time PCR assays with fluorescent-antibody assays for diagnosis of respiratory virus infections in children
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DOI：
10.1128/jcm.00216-06
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发表时间：
2006-07-01
影响因子：
9.4
通讯作者：
Morrow, Rhoda
中科院分区：
文献类型：
--
作者：
Kuypers, Jane;Wright, Nancy;Morrow, Rhoda
比较了常规荧光抗体（FA）方法与实时PCR检测方法在1年内收集的1，138份呼吸道疾病儿童标本中检测呼吸道合胞病毒（RSV）、流感病毒A型（FluA）、副流感病毒1型、2型和3型（PIV 1、PIV 2和PIV 3）、人偏肺病毒（MPV）和腺病毒（AdV）的情况。FA法检出至少一种病毒者436例（38.3%），PCR法检出至少一种病毒者608例（53.4%）（P < 0.001）。52份（4.6%）标本中FA的标本质量不足;其中13份（25%）经PCR检测呈阳性。相比之下，18例（1.6%）标本无法通过PCR进行分析;其中1例通过FA检测为阳性。在PCR和/或FA阳性的标本中，仅PCR阳性的标本数为18（7.0%）257例RSV，18例（13.4%）/134例FluA，25例（64.1%）39例接受PIV 1，8在所有标本中，MPV的检出率为6.6%，在FA阴性的702份标本中，MPV的检出率为9.5%。PCR和FA均阳性的标本中每毫升病毒拷贝数的平均值为6.7 × 10（7），显著高于仅PCR阳性的标本中的4.1 × 10（4）（p < 0.001）。PCR检测呼吸道病毒，尤其是副流感病毒和腺病毒的灵敏度明显高于FA检测。使用实时PCR鉴定儿童呼吸道病毒病原体将改善呼吸道疾病的诊断。
Conventional fluorescent-antibody (FA) methods were compared to real-time PCR assays for detection of respiratory syncytial virus (RSV), influenza virus type A (FluA), parainfluenza virus types 1, 2, and 3 (PIV1, PIV2, and PIV3), human metapneumovirus (MPV), and adenovirus (AdV) in 1,138 specimens from children with respiratory illnesses collected over a 1-year period. At least one virus was detected in 436 (38.3%) specimens by FA and in 608 (53.4%) specimens by PCR (P < 0.001). Specimen quality was inadequate for FA in 52 (4.6%) specimens; 13 of these (25%) were positive by PCR. In contrast, 18 (1.6%) specimens could not be analyzed by PCR; 1 of these was positive by FA. The number of specimens positive only by PCR among specimens positive by PCR and/or FA was 18 (7.0%) of 257 for RSV, 18 (13.4%) of 134 for FluA, 25 (64.1%) of 39 for PIV1, 8 (88.9%) of 9 for PIV2, 17 (30.1%) of 55 for PIV3, and 101 (76.5%) of 132 for AdV. MPV was detected in 6.6% of all specimens and in 9.5% of the 702 specimens negative by FA. The mean number of virus copies per milliliter in specimens positive by both PCR and FA was significantly higher, at 6.7 x 10(7), than that in specimens positive only by PCR, at 4.1 X 10(4) (p < 0.001). The PCR assays were significantly more sensitive than FA assays for detecting respiratory viruses, especially parainfluenza virus and adenovirus. Use of real-time PCR to identify viral respiratory pathogens in children will lead to improved diagnosis of respiratory illness.