Keywords
Antemortem Diagnosis
Polymerase Chain Reaction
How to Cite
Abstract
By using RT-PCR with primers targeted to the glycoprotein gene and RT-PCR and hemi-nested RT-PCR with primers targeted to the nucleoprotein gene, rabies viral RNA was identified in 95.2% of 21 samples, 18 of saliva and three of neck-skin biopsy, taken between hospitalization and death from a patient with clinical signs of rabies. Treatment administered to the patient included induction of coma and antiviral therapy. Each technique in isolation detected viral RNA in 90.5%, 57.1% and 85.7% of the samples, respectively. Our findings suggest that the parallel amplification of different regions of the genome of rabies virus can provide greater reliability to the antemortem diagnosis of rabies assisting medical decision regarding the application of the protocol treatment with antivirals.
References
King AMQ, Adams MJ, Carstens EB, Lefkowitz EJ. Virus taxonomy. Ninth report of the international Committee on Taxonomy of Viruses, San Diego, CA: Academic Press; 2012.
World Health Organization. WHO Expert Consultation on Rabies, First Report; 2005. (WHO Technical Report Series, 931).
Ministério da Saúde, Secretaria de Vigilância em Saúde. Controle, vigilância e profilaxia da raiva [monografia na internet]. Brasilia; 2010 [acesso em 29 dez. 2012]; Disponível em: http://portal.saude.gov.br/ portal/arquivos/pdf/prog_vig_raiva_dados_ ate_out_2010.pdf
Madhusudana SN, Sukumaran SM. Antemortem diagnosis and prevention of human rabies. Ann of Ind Acad of Neurol. 2008;11(1):3-12.
Franka R, Rupprecht CE. Treatment of rabies in the 21st century: curing the incurable? Future microbiol. 2011;6(10):1135-40.
Crepin P, Audry L, Rotivel Y, Gacoin A, Caroff C, Bourhy H. Intravitam diagnosis of human rabies by PCR using saliva and cerebrospinal fluid. J. clin microbiol. 1998; 36(4):1117-21.
David D, Rupprecht CE, Smith JS, Stram Y. Antemortem detection and virus characterization of three human rabies fatalities in Israel between 1996-1997. Isr. j. vet. med [periódico na internet]. 1999 [acesso em 29 dez. 2012];54(3). Disponível em: http://www.isrvma.org/ ImageToArticle/Files/Vol%2054%203.doc
Rupprecht CE, Cathleen AH, Hemachudha T. Rabies re-examined. Lancet. 2002;2: 327-43.
Trimarchi CV, Smith JS. Diagnostic evaluation. In: Jackson AC, Wunner WH, editores. Rabies. New York: Academic Press;2002. p. 307-49.
Smith J, Mcelhinney L, Parson G. Case report: rapid ante-mortem diagnosis of a human case of rabies imported into UK from Philippines. J. med. virol. 2003;69:1550-3.
Macedo CI, Carnieli Jr. P, Brandão PE, Rosa EST, Oliveira RN, Castilho JG et al. Diagnosis of human rabies cases by polymerase chain reaction of neck-skin samples. Braz. j. infect. dis. 2006; 10(5):341-5.
Wacharapluesadee S, Hemachudha T. Ante-and post-mortem diagnosis of rabies using nucleic acid-amplification tests. Expert rev. mol. diagn. 2010;10(2):207-18.
Dacheux L, Reynes JM, Buchy P, Sivuth O, Diop BM, Rousset D et al. A reliable diagnosis of human rabies based on analysis of skin biopsy specimens. Clin. Infect. dis. 2008;47(11):1410-7.
Willoughby RE Jr, Tieves KS, Hoffman GM, Ghanayem NS, Amlie-Lefond CM, Schwabe MJ et al. Survival after treatment of rabies with induction of coma. N. Engl. J. med. 2005;352:2508-14.
Ministério da Saúde, Secretaria de Vigilância em Saúde. Protocolo para tratamento da raiva humana no Brasil. Epidemiol. Serv. saúde. 2009;18(4):385-94.
Koprowski H. The mouse inoculation test. In: Meslin FX, Kaplan MM, Koprowski H, editores. Laboratory techniques in rabies. 4.ed. Geneva: World Health Organization; 1996. p. 80-7.
Orciari LA, Niezgoda M, Hanlon CA, Shaddock JH, Sanderlin DW, Yager PA et al. Rapid clearance of SAG-2 rabies virus from dogs after oral vaccination. Vaccine. 2001; 14(19):4511-8.
Sato G, Itou T, Miura Y, Mikami T, Ito M, Kurane I et al. Genetic and phylogenetic analysis of glycoprotein of RABV isolated from several species in Brazil. J. vet. Med. sci. 2004;66(7):747-53.
Hall TA. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl. Acids Symp. Ser. 1999;41:95-8.
Kumar S, Tamura K, Jakobsen IE, Nei M. MEGA 2: Molecular evolutionary genetic analysis software. Evolutionary Genetics Analysis Software, Bioinformatics. 2001;17(12):1244-5.
Wilde H. Failures of post-exposure rabies prophylaxis. Vaccine. 2007;25(44)7605-9.
Dean DJ, Abelseth MK, Atanasiu P. The fluorescent antibody test. In: Meslin FX, Kaplan MM, Koprowski H, editores. Laboratory techniques in rabies. 4.ed. Geneva: World Health Organization; 1996. p. 88-95.
Lembo T, Niezgoda M, Velasco-Villa A, Cleaveland S, Ernest E, Rupprecht CE. Evaluation of a direct, rapid immunohistochemical test for rabies diagnosis. Emerg. Infect. dis. 2006;12(2):310-3.
Bordignon J, Brasil-Dos-Anjos G, Bueno CR, Salvatiera-Oporto J, Dávila AM, Grisard EC et al. Detection and characterization of RABV in southern Brazil by PCR amplification and sequencing of the nucleoprotein gene. Arch. virol. 2005;150(4):695-708.
Franka R, Svrcek S, Madar M, Kolesarova M, Ondrejkova A, Ondrejka R et al. Quantification of the effectiveness of laboratory diagnostic of rabies using classical and molecular-genetic methods. Vet. Med. 2004;49(7):259-67.
Hemachudha T, Wacharapluesadee S. Antemortem diagnosis of human rabies. Clin. Infect. dis. 2004;39(7):1085-6.
Saengseesom W, Mitmoonpitak C, Kasempimolporn S, Sitprija V. Real-time PCR analysis of dog cerebrospinal fluid and saliva samples for ante-mortem diagnosis of rabies. Southeast Asian j. trop. med. public health. 2007;38(1):53-7.
Delmas O, Holmes EC, Talbi C, Larrous F, Dacheux L, Bouchier C et al. Genomic diversity and evolution of the lyssaviruses. PLos ONE. 2008;3(4):e2057.
Nagaraj T, Vasanth JP, Desai A, Kamat A, Madhusudana SN, Ravi V. Ante mortem diagnosis of human rabies using saliva samples: comparison of real time and conventional RT-PCR techniques. J. clin. virol. 2006;36(1):17-23.
Carnieli P Jr, Fahl WO, Brandão PE, Oliveira R de N, Macedo CI, Durymanova E et al. Comparative analysis of rabies virus isolates from Brazilian canids and bats based on the G gene and G-L intergenic region. Arch. virol. 2010;155(6):941-8.
Sato G, Tanabe H, Shoji Y, Takuya I, Ito FH, Sato T et al. Rapid discrimination of rabies viruses isolated from various host species in Brazil by multiplex reverse transcriptionpolymerase chain reaction. J. clin. virol. 2005;33(4):267-73.
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Copyright (c) 2013 Juliana Galera Castilho, Carla Isabel Macedo, Saulo de Oliveira Santos, Rafael de Novaes Oliveira, Samira Maria Achkar, Maria Luiza Carrieri, Ivanete Kotait, Pedro Carnieli Jr