Using the LN34 Pan-Lyssavirus Real-Time RT-PCR Assay for Rabies Diagnosis and Rapid Genetic Typing from Formalin-Fixed Human Brain Tissue

被引:9
作者
Condori, Rene Edgar [1 ]
Niezgoda, Michael [1 ]
Lopez, Griselda [2 ]
Acosta Matos, Carmen [3 ]
Diaz Mateo, Elinna [4 ]
Gigante, Crystal [1 ,5 ]
Hartloge, Claire [1 ,5 ]
Pereira Filpo, Altagracia [2 ]
Haim, Joseph [6 ]
Satheshkumar, Panayampalli Subbian [1 ]
Petersen, Brett [1 ]
Wallace, Ryan [1 ]
Olson, Victoria [1 ]
Li, Yu [1 ]
机构
[1] Ctr Dis Control & Prevent, Pox & Rabies Branch, Atlanta, GA 30329 USA
[2] Lab Salud Publ Dr Defillo, Santo Domingo 10105, Dominican Rep
[3] Direcc Salud Pedernales, Pedernales 84000, Dominican Rep
[4] Ctr Prevenc & Control Enfermedades Transmitidas V, Santo Domingo 10308, Dominican Rep
[5] Oak Ridge Inst Sci & Educ, Oak Ridge, TN 37830 USA
[6] Minist Agr Resources Nat & Dev Rural, Dept Anim Hlth, Ht 6110, Haiti
来源
VIRUSES-BASEL | 2020年 / 12卷 / 01期
关键词
rabies virus; lyssavirus; diagnosis; LN34; assay; formalin-fixed tissue; genetic typing; ANTIGEN; DOG; PREVENTION; SYSTEM; VIRUS;
D O I
10.3390/v12010120
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Human rabies post mortem diagnostic samples are often preserved in formalin. While immunohistochemistry (IHC) has been routinely used for rabies antigen detection in formalin-fixed tissue, the formalin fixation process causes nucleic acid fragmentation that may affect PCR amplification. This study reports the diagnosis of rabies in an individual from the Dominican Republic using both IHC and the LN34 pan-lyssavirus real-time RT-PCR assay on formalin-fixed brain tissue. The LN34 assay generates a 165 bp amplicon and demonstrated higher sensitivity than traditional PCR. Multiple efforts to amplify nucleic acid fragments larger than 300 bp using conventional PCR were unsuccessful, probably due to RNA fragmentation. Sequences generated from the LN34 amplicon linked the case to the rabies virus (RABV) strain circulating in the Ouest Department of Haiti to the border region between Haiti and the Dominican Republic. Direct sequencing of the LN34 amplicon allowed rapid and low-cost rabies genetic typing.
引用
收藏
页数:11
相关论文
共 35 条
  • [1] [Anonymous], Rabies
  • [2] Evaluation of formalin-free tissue fixation for RNA and microRNA studies
    Arzt, Lisa
    Kothmaier, Hannelore
    Quehenberger, Franz
    Halbwedl, Iris
    Wagner, Karin
    Maierhofer, Theresa
    Popper, Helmut H.
    [J]. EXPERIMENTAL AND MOLECULAR PATHOLOGY, 2011, 91 (02) : 490 - 495
  • [3] Control and prevention of canine rabies: The need for building laboratory-based surveillance capacity
    Banyard, Ashley C.
    Horton, Daniel L.
    Freuling, Conrad
    Mueller, Thomas
    Fooks, Anthony R.
    [J]. ANTIVIRAL RESEARCH, 2013, 98 (03) : 357 - 364
  • [4] Overview of rabies in the Americas
    Belotto, A
    Leanes, LF
    Schneider, MC
    Tamayo, H
    Correa, E
    [J]. VIRUS RESEARCH, 2005, 111 (01) : 5 - 12
  • [5] Distribution of rabies antigen in infected brain material: determining the reliability of different regions of the brain for the rabies fluorescent antibody test
    Bingham, J
    van der Merwe, M
    [J]. JOURNAL OF VIROLOGICAL METHODS, 2002, 101 (1-2) : 85 - 94
  • [6] Virology, Immunology and Pathology of Human Rabies During Treatment
    Caicedo, Yolanda
    Paez, Andres
    Kuzmin, Ivan
    Niezgoda, Michael
    Orciari, Lillian A.
    Yager, Pamela A.
    Recuenco, Sergio
    Franka, Richard
    Velasco-Villa, Andres
    Willoughby, Rodney E., Jr.
    [J]. PEDIATRIC INFECTIOUS DISEASE JOURNAL, 2015, 34 (05) : 520 - 528
  • [7] Chacko Kadavil, 2016, Qatar Med J, V2016, P15, DOI 10.5339/qmj.2016.15
  • [8] Coertse J, 2011, J S AFR VET ASSOC, V82, P250
  • [9] A Systematic Review of Human Bat Rabies Virus Variant Cases: Evaluating Unprotected Physical Contact with Claws and Teeth in Support of Accurate Risk Assessments
    Dato, Virginia M.
    Campagnolo, Enzo R.
    Long, Jonah
    Rupprecht, Charles E.
    [J]. PLOS ONE, 2016, 11 (07):
  • [10] Dean D. J., 1996, LAB TECHNIQUES RABIE, P88