摘要
目的探讨多重实时荧光定量PCR(MRT-PCR)检测9种常见呼吸道病原体的临床应用价值。方法采用Primer Express3.0(ABI)和Primer3 Input 4.0设计引物和探针建立MRT-PCR法,通过构建质粒标准品分析该方法的最低检出限和特异性。并对204例临床标本中副流感病毒1、2、3型、肺炎支原体、肺炎衣原体、呼吸道合胞病毒、腺病毒、甲型流感病毒、乙型流感病毒进行回顾性检测。结果 MRT-PCR法检测病原体的最低检出限达103copies/m L,特异性达100%,无交叉反应。204例咽拭子标本中,MRT-PCR检出呼吸道合胞病毒23例,副流感病毒1、2、3型13例,腺病毒15例,肺炎支原体15例,肺炎衣原体3例,甲型流感病毒13例与乙型流感病毒6例。该结果与其他试剂报告结果完全一致。结论多重实时荧光定量PCR具有快速、准确、特异性强等特点,在呼吸道病原体检测方面有重要价值。
Objective To explore the clinical significance of nine common respiratory pathogens detected by multiple real-time quantitative PCR（ MRT-PCR） assay. Methods The primers and probes for respiratory viruses were designed with Primer Express 3. 0（ ABI） and Primer3 Input 4. 0. The lowest detectable limit and specificity of MRT-PCR were analyzed by plasmid standard constructing. Parainfluenza type 1,2,3,mycoplasma,chlamydia,respiratory syncytial virus,adenovirus,influenza virus A and B in 204 throat swabs were retrospectively detected by the established method. Results The lowest detectable limit of MRT-PCR for seven pathogens was 103 copies / ml,and its specificity was 100%. Furthermore,there was no cross reaction. 204 throat swabs were detected by MRT-PCR,including respiratory syncytial virus（ 23 cases）,parainfluenza virus type 1,2,3（ 13cases）,adenovirus（ 15 cases）,mycoplasma pneumoniae（ 15 cases）,chlamydia pneumoniae（ 3 cases）,influenza virus A（ 13 cases） and influenza virus B（ 6 cases）. The results in this study were consistent with the results determined by other methods. Conclusion The multiplex real-time quantitative PCR is characterized by rapidity,accuracy,and specificity,which has great value in detection of respiratory pathogens.
出处
《标记免疫分析与临床》
CAS
2015年第11期1160-1164,共5页
Labeled Immunoassays and Clinical Medicine
