首页> 美国卫生研究院文献>Journal of Clinical Microbiology >Rapid diagnosis of human parainfluenza virus type 1 infection by quantitative reverse transcription-PCR-enzyme hybridization assay.
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Rapid diagnosis of human parainfluenza virus type 1 infection by quantitative reverse transcription-PCR-enzyme hybridization assay.

机译:通过定量逆转录-PCR-酶杂交测定法快速诊断人副流感病毒1型感染。

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摘要

The detection and quantitation of human parainfluenza virus type 1 (HPIV-1) RNA in nasal wash specimens from 49 children with lower respiratory infections were performed by a reverse transcription-PCR-enzyme hybridization assay (RT-PCR-EHA). The HPIV-1 RT-PCR-EHA was then used to test 40 samples from asymptomatic children. Primers and probes were designed from regions within the HPIV-1 hemagglutinin-neuraminidase gene which are highly conserved among all known genotypes. HPIV-1 was detected in all nine children who were culture positive. Other common respiratory viruses (HPIV-2, -3, and -4, mumps virus, respiratory syncytial virus, and influenza virus) were not detected by the HPIV-1 assay. Forty symptomatic children were negative by culture, and four of these were positive by RT-PCR-EHA. All of the samples from asymptomatic children were negative by culture and RT-PCR-EHA. RT-PCR-EHA was 100% sensitive (95% confidence interval, 0.66 to 1.00) and 95% specific (95% confidence interval, 0.88 to 0.99) compared with culture. The four false-positive results (relative to the results of culture) were in children with lower respiratory infections compatible with HPIV-1 infection and suggest that RT-PCR-EHA may be more sensitive than culture. Our data indicate that HPIV-1 may be underdiagnosed by routine culturing methods. RT-PCR-EHA has been demonstrated to be an easy, rapid, sensitive, and specific test for diagnosing HPIV-1 infection and provides a methodology for the rapid detection of closely related respiratory viruses.
机译:通过逆转录-PCR-酶杂交试验(RT-PCR-EHA)对49名下呼吸道感染患儿的鼻洗样本中的人副流感病毒1型(HPIV-1)RNA进行检测和定量。然后,使用HPIV-1 RT-PCR-EHA测试来自无症状儿童的40个样品。从HPIV-1血凝素神经氨酸酶基因内的区域设计引物和探针,这些区域在所有已知基因型中高度保守。在培养阳性的所有9名儿童中均检测到HPIV-1。 HPIV-1分析未检测到其他常见的呼吸道病毒(HPIV-2,-3和-4,腮腺炎病毒,呼吸道合胞病毒和流感病毒)。有症状的40名儿童在培养中为阴性,其中有4名通过RT-PCR-EHA阳性。通过培养和RT-PCR-EHA,所有无症状儿童的样品均为阴性。与培养相比,RT-PCR-EHA的敏感性为100%(95%置信区间,0.66至1.00)和95%特异性(95%置信区间,0.88至0.99)。四个假阳性结果(相对于培养结果)是在下呼吸道感染与HPIV-1感染相容的患儿中,提示RT-PCR-EHA可能比培养更敏感。我们的数据表明,HPIV-1可能无法通过常规培养方法进行诊断。 RT-PCR-EHA已被证明是诊断HPIV-1感染的简便,快速,灵敏和特异的检测方法,它为快速检测紧密相关的呼吸道病毒提供了一种方法。

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