CAJ | 학술논문

目的人副流感病毒1,2,3型是呼吸道感染的主要病原.本研究建立了特异、快速、灵敏的多重荧光定量RT-PCR方法用于人副流感1,2,3型病毒临床标本检测.方法针对人副流感1,2,3型病毒设计特异性引物探针,优化荧光RT-PCR反应条件.应用体外转录方法分别制备人副流感1,2,3型病毒的标准品.验证荧光定量RT-PCR方法的特异性,敏感性和稳定性.结果该方法对人副流感1,2,3型病毒核酸检测有高度特异性,检测的灵敏度HPIV1为10个拷贝,HPIV2为100个拷贝,HPIV3为100个拷贝.可从临床患者鼻咽吸出物标本中直接检出.结论本研究建立的LNA探针同时检测人副流感病毒1,2,3型多重荧光定量RT-PCR方法具有较高的特异性和敏感性.适用于临床早期诊断和实验室病原谱筛查.
목적 인부류감병독1,2,3형시호흡도감염적주요병원.본연구건립료특이、쾌속、령민적다중형광정량RT-PCR방법용우인부류감1,2,3형병독림상표본검측.방법 침대인부류감1,2,3형병독설계특이성인물탐침,우화형광RT-PCR반응조건.응용체외전록방법분별제비인부류감1,2,3형병독적표준품.험증형광정량RT-PCR방법적특이성,민감성화은정성.결과 해방법대인부류감1,2,3형병독핵산검측유고도특이성,검측적령민도HPIV1위10개고패,HPIV2위100개고패,HPIV3위100개고패.가종림상환자비인흡출물표본중직접검출.결론 본연구건립적LNA탐침동시검측인부류감병독1,2,3형다중형광정량RT-PCR방법구유교고적특이성화민감성.괄용우림상조기진단화실험실병원보사사.
Objective Human parainfluenza virus (HPIV) types 1,2 and 3 are major viral pathogens responsible for upper and lower respiratory tract infections.In this study,a real-time RT-PCR was developed using multiplex primers-probe (HPIV-1,2,3) for the simultaneous detection of both HPIV1,HPIV2 and HPIV3 genomes.Methods Optimal primers and probes were designed using specialized software.The conditions for multiplex real-time RT-PCR had been optimized.The synthesis of RNA standards of HPIV1,2,3 were used a T7 RNA polymerase.Check the specificity sensitivities and stability of one step RT-PCR assay.Results Obtained in a 10-fold dilution series assay demonstrate a high sensitivity of the assay with a lowest detection limit of 10 copies for HPIV1,100 copies for HPIV2 and 100 copies for HPIV3.Conclusion The assays demonstrates an improved sensitivity and scope of detecting HPIV1,2,3 viruses relative to routine antigen detection assays while the quantitative utility may facilitate investigation of the pre-diagnosis and respiratory virus pathogenesis.