Comparison of Automated Microarray Detection with Real-Time PCR Assays for Detection of Respiratory Viruses in Specimens Obtained from Children
Comparison of Automated Microarray Detection with Real-Time PCR Assays for Detection of Respiratory Viruses in Specimens Obtained from Children
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DOI：
10.1128/jcm.01297-08
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发表时间：
2009-03-01
影响因子：
9.4
通讯作者：
Corbeil, Jacques
中科院分区：
文献类型：
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作者：
Raymond, Frederic;Carbonneau, Julie;Corbeil, Jacques
呼吸道病毒感染是一个主要的健康问题，也是幼儿接受检测、咨询和住院治疗的主要原因。我们开发并比较了两种检测方法，可检测多达23种经常感染儿童的不同呼吸道病毒。第一种方法包括96孔板格式的TaqMan实时定量PCR检测。第二次包括多重PCR，随后引物延伸和微阵列杂交在一个集成的分子诊断设备，Infiniti分析仪。两种检测方法均可检测A、B、C和E组腺病毒；冠状病毒HKU1、229E、NL63和OC43；肠病毒A、B、C和D；A、B基因型鼻病毒；甲型和乙型流感病毒；人偏肺病毒（HMPV） A、B、人呼吸道合胞病毒（HRSV） A、B；还有1、2、3型副流感病毒。这些试验用于鉴定从因呼吸道感染住院的儿童获得的221份鼻咽吸入物中的病毒。221份标本中有81.4%的标本至少用两种方法中的一种检测出呼吸道病毒：HRSV A阳性率为10.0%，HRSV B阳性率为38.0%，流感病毒A阳性率为13.1%，冠状病毒阳性率为8.6%，鼻病毒或肠病毒阳性率为13.1%，腺病毒阳性率为7.2%，HMPV阳性率为4.1%，副流感病毒阳性率为1.5%。13.1%的标本存在多重病毒感染。两种方法在94.1%的标本中结果一致。这些检测可以对医院环境中感染儿童的呼吸道病毒进行彻底的病原学评估，并有助于公共卫生干预。
Respiratory virus infections are a major health concern and represent the primary cause of testing consultation and hospitalization for young children. We developed and compared two assays that allow the detection of up to 23 different respiratory viruses that frequently infect children. The first method consisted of single TaqMan quantitative real-time PCR assays in a 96-well-plate format. The second consisted of a multiplex PCR followed by primer extension and microarray hybridization in an integrated molecular diagnostic device, the Infiniti analyzer. Both of our assays can detect adenoviruses of groups A, B, C, and E; coronaviruses HKU1, 229E, NL63, and OC43; enteroviruses A, B, C, and D; rhinoviruses of genotypes A and B; influenza viruses A and B; human metapneumoviruses (HMPV) A and B, human respiratory syncytial viruses (HRSV) A and B; and parainfluenza viruses of types 1, 2, and 3. These tests were used to identify viruses in 221 nasopharyngeal aspirates obtained from children hospitalized for respiratory tract infections. Respiratory viruses were detected with at least one of the two methods in 81.4% of the 221 specimens: 10.0% were positive for HRSV A, 38.0% for HRSV B, 13.1% for influenzavirus A, 8.6% for any coronaviruses, 13.1% for rhinoviruses or enteroviruses, 7.2% for adenoviruses, 4.1% for HMPV, and 1.5% for parainfluenzaviruses. Multiple viral infections were found in 13.1% of the specimens. The two methods yielded concordant results for 94.1% of specimens. These tests allowed a thorough etiological assessment of respiratory viruses infecting children in hospital settings and would assist public health interventions.