| 1 | Development of a multiplex one-step real-time RT-PCR assay for the simultaneous detection of eight viruses associated with febrile rash illnesses显示文摘Fever and rash illnesses(FRIs)are a series of common diseaseswith fever and rashes as clinicalmanifestations,most of which are caused by viral infection.The rashes of FRIs are generally nonspecific;therefore it is difficult to identify FRIassociated viruses solely based on clinical symptoms.To achieve rapid and accurate identification of FRI pathogens,a multiplex one-step real-time reverse transcription-polymerase chain reaction(RT-PCR)assay was developed and evaluated in this study.Primers and probes were selected for the detection of measles virus(MeV),rubella virus(RV),human enterovirus(EV),varicella-zoster virus(VZV),dengue virus(DENV),human parvovirus B19(B19),Epstein-Barr virus(EBV),and human herpes virus 6(HHV-6),which cover the most common pathogenic viruses of FRIs.Detection of the eight FRI-associated viruses,which was divided into two groups/tubes,was simultaneously performed under universal optimized reaction conditions in multiplex one-step real-time RT-PCR assay.The multiplex realtime RT-PCR showed high sensitivity and specificity in detecting the eight FRI-associated viruses.The limits of detection(LODs)for the eight viruses were in the range of 47–177 copies/reaction,and no cross reactions for the eight FRIassociated viruses were found in the multiplex assay.In addition,the results of the multiplex real-time RT-PCR assay were consistent with the results of a monoplex real-time RT-PCR assay and sequencing for clinical specimens obtained from FRI patients.With its advantages of high efficiency and rapid and accurate diagnosis,multiplex real-time RT-PCR was very feasible for the early diagnosis of FRI pathogenic viruses and would be of great help for the proper treatment,monitoring,and initiation of preventive measures for FRI cases. | Aili Cui Shulei Wang Qiang Zhang Huiling Wang Zhen Zhu Aqian Li Qinqin Song Yanzhe Hao Jilan He Wenbo Xu Yan Zhang | 2020 | Biosafety and Health2020，2，2： | 5 |
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