Rapid Detection of Filoviruses by Real-time TaqMan Polymerase Chain Reaction Assays

• Published in: Virologica Sinica Vol. 27; no. 5; pp. 273 - 277
• Main Authors: Huang, Yi, Wei, Hongping, Wang, Yunpeng, Shi, Zhengli, Raoul, Herve, Yuan, Zhiming
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer-Verlag 01-10-2012; SP Wuhan Institute of Virology, CAS; State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China%Laboratoire P4 Jean Mérieux, National Institute for Health and Medical Research(Inserm), 69365 Lyon Cedex 07, France
• Subjects: Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; China; Clinical Laboratory Techniques - methods; Data processing; Differential diagnosis; Ebola virus; Ebolavirus - genetics; Ebolavirus - isolation & purification; fever; Filovirus; genes; Hemorrhagic fever; Hemorrhagic Fever, Ebola - diagnosis; Hemorrhagic Fever, Ebola - veterinary; Humans; Marburg virus; Marburg Virus Disease - diagnosis; Marburgvirus - genetics; Marburgvirus - isolation & purification; Medical Microbiology; messenger RNA; Microbial Genetics and Genomics; Microbiology; monitoring; Mortality; Nucleoproteins; Nucleoproteins - genetics; Oncology; Polymerase chain reaction; Primates; Quarantine; Real-Time Polymerase Chain Reaction - methods; Research Article; Ribonucleoproteins - genetics; RNA; RT-PCR分析; Sensitivity and Specificity; TaqMan; Transcription; Vaccines; Viral Core Proteins - genetics; Viral Proteins - genetics; virion; Virions; Virology; Virology - methods; viruses; Zaire Ebola virus; 丝状; 埃博拉病毒; 实时; 快速检测; 荧光定量聚合酶链反应; 诊断测试; China; Ebola virus; Nucleoprotein (NP) gene; Marburg virus; TaqMan RT-PCR
• ISSN: 1674-0769; 1995-820X
• DOI: 10.1007/s12250-012-3252-y

Ebola virus （EBOV） and Marburg virus （MARV） are causative agents of severe hemorrhagic fever with high mortality rates in humans and non-human primates and there is currently no licensed vaccine or therapeutics. To date, there is no specific laboratory diagnostic test in China, while there is a national need to provide differential diagnosis during outbreaks and for instituting acceptable quarantine procedures. In this study, the TaqMan RT-PCR assays targeting the nucleoprotein genes of the Zaire Ebolavirus （ZEBOV） and MARV were developed and their sensitivities and specificities were investigated. Our results indicated that the assays were able to make reliable diagnosis over a wide range of virus copies from 103 to 109, corresponding to the threshold of a standard RNA transcript. The results showed that there were about 101 RNA copies per milliliter of virus culture supernatant, equivalent to 10,000 RNA molecules per infectious virion, suggesting the presence of many non-infectious particles. These data indicated that the TaqMan RT-PCR assays developed in this study will be suitable