title: Quantum dot-fluorescence in situ hybridisation for Ectromelia virus detection based on biotin-streptavidin interactions.
abstract: Ectromelia virus (ECTV) is an pathogen that can lead to a lethal, acute toxic disease known as mousepox in mice. Prevention and control of ECTV infection requires the establishment of a rapid and sensitive diagnostic system for detecting the virus. In the present study, we developed a method of quantum-dot-fluorescence based in situ hybridisation for detecting ECTV genome DNA. Using biotin-dUTP to replace dTTP, biotin was incorporated into a DNA probe during polymerase chain reaction. High sensitivity and specificity of ECTV DNA detection were displayed by fluorescent quantum dots based on biotin-streptavidin interactions. ECTV DNA was then detected by streptavidin-conjugated quantum dots that bound the biotin-labelled probe. Results indicated that the established method can visualise ECTV genomic DNA in both infected cells and mouse tissues. To our knowledge, this is the first study reporting quantum-dot-fluorescence based in situ hybridisation for the detection of viral nucleic acids, providing a reference for the identification and detection of other viruses.
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Zheng
authors.affiliation: National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China. Electronic address: zhangxe@ibp.ac.cn.
authors.forename: Xian-En
authors.initials: XE
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. Electronic address: wanghz@wh.iov.cn.
authors.forename: Hanzhong
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D001710:Biotin / Q000737:chemistry*; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000032:analysis*; D004481:Ectromelia virus / Q000235:genetics* / Q000648:ultrastructure; D004482:Ectromelia, Infectious / Q000821:virology; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D046529:Microscopy, Electron, Transmission; D045663:Quantum Dots / Q000737:chemistry*; D013154:Spleen / Q000648:ultrastructure / Q000821:virology; D019809:Streptavidin / Q000737:chemistry*; D013623:Tail / Q000821:virology; D014709:Vero Cells
chemical_list: D004279:DNA, Viral; D001710:Biotin; D019809:Streptavidin
keywords: Ectromelia virus; Fluorescence in situ hybridisation; Quantum-dot; Virus DNA detection