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呼吸道合胞病毒实时荧光定量pcr检测

, PP. 847-852

Keywords: 呼吸道合胞病毒,实时监测,荧光定量pcr,taqman

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Abstract:

目的建立一种快速、准确诊断人呼吸道合胞病毒(hrsv)早期感染的实时荧光定量pcr检测方法。方法以hrsv最保守的n基因序列为参考,设计两对特异的引物和一条taqman荧光探针,在lightcycler定量pcr检测仪上对93例下呼吸道感染患儿进行hrsvrna检测,并与病毒分离培养、常规pcr、巢式pcr方法和elisa法相比较。结果以pcr模板浓度来定义,该方法线形范围为1×102~1×107cdnacopies/μl,灵敏度达1×102cdnacopies/μl,和巢式pcr相同,比常规pcr高10倍。用人脊髓灰质炎病毒ⅰ型、柯萨奇病毒2型、甲型、乙型流感病毒、腺病毒7型作对照,均无预期扩增产物和荧光的产生;该方法在lightcycler和rotor-gene两种仪器上的定量结果具有一致性;采用该法对93例患儿行fq-pcr检测出阳性44例(43.9%),elisa方法检测阳性4例(4.3%),两者相关性不高。hrsv浓度的高低与患儿临床症状之间的关系不是很明显。结论该方法可快速、灵敏、特异、定量检测hrsv,在hrsv感染的早期诊断和疗效监测方面具有良好的应用前景。

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