实时 RT-PCR、拭子瓶培养和常规细胞培养在检测住院患者中流行的甲型流感（H1N1）中的比较。
Comparison of real-time RT-PCR, shell vial culture, and conventional cell culture for the detection of the pandemic influenza A (H1N1) in hospitalized patients.
机构信息
Clinical Microbiology and Infectious Diseases Department, Hospital General Universitario Gregorio Marañon, CIBER de Enfermedades Respiratorias (CIBERES), Madrid, Spain.
出版信息
Diagn Microbiol Infect Dis. 2011 Apr;69(4):428-31. doi: 10.1016/j.diagmicrobio.2010.11.007.
The emergence of the pandemic influenza virus A H1N1 has made fast and accurate diagnosis essential. However, few well-validated diagnostic techniques exist. The real-time RT-PCR developed by the Centers for Disease Control and Prevention (CDC) is the recommended technique. Our objective was to compare the CDC real-time RT-PCR assay, shell vial (SV), and conventional cell culture [with Madine-Darby canine kidney (MDCK) cells and A549] for the detection of pandemic influenza A H1N1 in hospitalized patients. We performed a prospective study comparing the efficacy of 5 diagnostic techniques (RTPCR, SV in A549, SV in MDCK, conventional cell culture in A549, and conventional cell culture in MDCK) using nasopharyngeal swabs from patients ≥18 years of age hospitalized with clinical symptoms of influenza at our institution. Detection of the virus by conventional culture was considered the gold standard. An "extended gold standard" was also used to recalculate validity values. The sensitivities, specificities, positive predictive values, and negative predictive values (NPVs) for the detection of influenza A H1N1, determined using conventional culture as the gold standard, were, respectively, as follows: RT-PCR: 95.6, 82.3, 78.3, 96.5%; SVA549: 91.2, 99.01, 98.4, 94.4%; SV-MDCK: 82.3, 100, 100, 89.4%; tube-A549: 94.12, 100, 100, 96.2%; tube-MDCK: 86.7, 100, 100, 91.9%. Sensitivities and NPVs using an extended gold standard were as follows: RT-PCR: 96.5%, 96.6%; SV-A549: 73.3%, 78.5%; SV-MDCK: 65.1%, 73.7%; tube-A549: 74.4%, 79.2%; tube-MDCK: 68.6%, 75.7%. The average time to detect pandemic influenza A H1N1 by RT-PCR, SV culture, and conventional culture was, respectively, 4 h, 48 h, and 7 days. Real-time RT-PCR displayed high sensitivity and specificity for the detection of influenza A H1N1 in adult patients when compared with conventional techniques. In addition, the A549 cell line was not inferior to the MDCK line.
甲型 H1N1 大流行流感病毒的出现使得快速准确的诊断变得至关重要。然而，目前可用的验证良好的诊断技术却很少。疾病预防控制中心（CDC）开发的实时 RT-PCR 是推荐使用的技术。我们的目的是比较 CDC 实时 RT-PCR 检测法、细胞瓶（SV）和常规细胞培养[使用 Madine-Darby 犬肾（MDCK）细胞和 A549 细胞]在住院患者中检测大流行流感 A H1N1 的效果。我们进行了一项前瞻性研究，比较了 5 种诊断技术（RT-PCR、SV 培养于 A549 细胞、SV 培养于 MDCK 细胞、常规细胞培养于 A549 细胞和常规细胞培养于 MDCK 细胞）在我院因流感临床症状住院的≥18 岁患者鼻拭子中的疗效。将常规培养检测到病毒作为金标准。还使用了“扩展金标准”重新计算了有效性值。使用常规培养作为金标准时，甲型 H1N1 检测的灵敏度、特异性、阳性预测值和阴性预测值（NPV）分别为：RT-PCR：95.6%、82.3%、78.3%、96.5%；SV-A549：91.2%、99.01%、98.4%、94.4%；SV-MDCK：82.3%、100%、100%、89.4%；管-A549：94.12%、100%、100%、96.2%；管-MDCK：86.7%、100%、100%、91.9%。使用扩展金标准时的灵敏度和 NPV 分别为：RT-PCR：96.5%、96.6%；SV-A549：73.3%、78.5%；SV-MDCK：65.1%、73.7%；管-A549：74.4%、79.2%；管-MDCK：68.6%、75.7%。甲型 H1N1 大流行流感病毒的平均检测时间分别为 RT-PCR 4 小时、SV 培养 48 小时和常规培养 7 天。实时 RT-PCR 与常规技术相比，对成人患者甲型 H1N1 的检测具有较高的灵敏度和特异性。此外，A549 细胞系并不逊于 MDCK 细胞系。
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