Simultaneous detection of human parainfluenza viruses 1, 2, 3 by multiplex real-time reverse transcription-polymerase chain reaction with LNA probes
10.3760/cma.j.issn.1003-9279.2012.05.020
- VernacularTitle: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方法的建立
- Author:
Yi-Xin JI
1
;
Nai-Ying MAO
;
Huan-Huan WANG
;
Zheng-De XIE
;
Wen-Bo XU
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Parainfluenza virus Ⅰ,human;
Parainfluenza virus Ⅱ,human;
Parainfluenza virus Ⅲ,human;
Polymerase chain reaction;
DNA probes,LNA
- From:
Chinese Journal of Experimental and Clinical Virology
2012;26(5):388-390
- CountryChina
- Language:Chinese
-
Abstract:
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.
