Discrimination between dog-related and vampire bat-related rabies viruses in Brazil by strain-specific reverse transcriptase-polymerase chain reaction and restriction fragment length polymorphism analysis

Ito, Makoto; Itou, Takuya; Shoji, Youko; Sakai, Takeo; Ito, Fumio; Arai, Yohko T.; Takasaki, Tomohiko; Kurane, Ichiro

doi:10.1016/s1386-6532(02)00048-3

Cited by 49 publications

(38 citation statements)

References 32 publications

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“…Rabies viruses (RVs) isolated in Brazil belong to genotype 1 of genus lyssavirus, family Rhabdoviridae. Ito et al [7,8] reported that main reservoirs of RVs were dogs and vampire bats and that the RVs were classified into dogrelated RVs and vampire bat-related RVs in Brazil. In some areas of Brazil, the rabies outbreaks by both dog-and batrelated RVs occurred at the same time [7].…”

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Molecular Epidemiological Analysis of Bat Rabies Viruses in Brazil

Kobayashi

Sato

Shoji

et al. 2005

The Journal of Veterinary Medical Science

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ABSTRACT. A molecular epidemiological analysis was performed in 19 rabies viruses (RVs) isolated from haematophagous, frugivorous and insectivorous bats, in Sao Paulo, Brazil. The authors carried out RT-PCR for amplification of the RV nucleoprotein (N) gene, and determined 1,335 nucleotide sequences of N gene by direct sequencing method. Phylogenetic analysis, which was based on the N gene of Brazilian RV isolates identified presently and previously, revealed that RVs isolated from bats were genetically divided into four lineages had a tendency to depend on the host bat species. Rabies viruses (RVs) isolated in Brazil belong to genotype 1 of genus lyssavirus, family Rhabdoviridae. Ito et al. [7,8] reported that main reservoirs of RVs were dogs and vampire bats and that the RVs were classified into dogrelated RVs and vampire bat-related RVs in Brazil. In some areas of Brazil, the rabies outbreaks by both dog-and batrelated RVs occurred at the same time [7]. RVs have been isolated from not only haematophagous bats but also frugivorous and insectivorous bats in Brazil. In Argentina, RVs isolated from frugivorous bats (Artibeus lituratus) were identified to be the variant of RV associated with haematophagous bats by use of the monoclonal antibody panels [4]. Recently we have demonstrated that RVs isolated from Brazilian frugivorous bats (Artibeus spp.) were phylogenetically characterized as the vampire bat-related RV [18].There are many reports of human cases on infection with insectivorous bat RVs, and the number of the cases tend to increase in North America [10][11][12]. In 2002, Favi et al. [5] reported that insectivorous bat (Tadarida brasiliensis) RV variant was isolated from a human patient in Chile. In Colombia and Venezuela insectivorous bat RV variants were also isolated from dogs and cats, which were main RVtransmitters to human [2,14]. Although some kinds of insectivorous bats have been diagnosed as RV-positive in Brazil, there have been no reports of the human case on infection with insectivorous bat RV variants until now. RVpositive insectivorous bats were, however, found in urban areas, and this fact has been considered as important problems in public health [1,6,17,21].As there is little information about the genetic characteristics of bat-related RVs in Brazil, authors determined nucleotide sequences on the N gene of RVs isolated from 9 Brazilian bat species. Phylogenetic analysis was also performed for the epidemiological study with the isolates identified previously. MATERIALS AND METHODS Viruses:In this study, 19 RVs were isolated from 3 haematophagous bats (Desmodus rotundus), 1 frugivorous bat (Artibeus lituratus), 13 insectivorous bats (3 Nyctinomops laticaudatus, 2 Eumops auripendulus, 4 Eptesicus furinalis, 2 Molossus molossus, 1 Molossus abrasus and 1 Molossus rufus) and 2 unclassified bats, and their nucleoprotein gene sequences were determined. Brain materials from these bats were diagnosed as RV-positive by immunofluorescent antibody test (FAT) using a rabbit polyclonal anti-rabies nucle...

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Molecular Epidemiological Analysis of Bat Rabies Viruses in Brazil

Kobayashi

Sato

Shoji

et al. 2005

The Journal of Veterinary Medical Science

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“…RNAs were extracted from mouse brain suspensions with a commercial reagent (QIAamp Viral RNA Mini Kit, QIAGEN). RT-PCR and sequencing of the complete coding region of the nucleoprotein (N) gene was performed using sense primers: JW12 (5'-ATGTAACACCYCTACAAT-3', [4]), P1 (5'-CTA-CAATGGATGCCGACAAGA-3', [5]), BRABN-S1 (5'-GGACTAGCTATGGAATCCTG -3', [6]) and BRABN-S3 (5'-GGACTGGTGTCATTTACAGG-3', [6]), and antisense primers: N8 (5'-AGTTTCTTCAGCCATCTC-3', [7]), P2 (5'-CCCATATAACATCCAACAAAGTG-3', [5]) and BRABN-C3 (5'-TGTCCAGAGATTTTGCTCA-3'). The sequences of 1,332 bp corresponding to nucleotides 89-1,420 on the N gene determined in this study and retrieved from GenBank (Table 1) were used for phylogenetic analy-sis by the neighbor-joining methods using the Clustal X program [13] and the bootstrap probabilities of each node were calculated using 1,000 replicates.…”

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Genetic Characterization of Rabies Viruses Isolated from Frugivorous Bat (Artibeus spp.) in Brazil

Shoji

Kobayashi

Sato

et al. 2004

The Journal of Veterinary Medical Science

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ABSTRACT. In Latin America, rabies cases related to frugivorous bats have been reported since 1930's. Recently, two viruses isolated from Artibeus lituratus were proved to be vampire bat variants by monoclonal antibodies panels [2], but their genetic information is not well known. In this report, four rabies viruses were isolated from frugivorous bats (Artibeus spp.) in Brazil and their nucleoprotein gene sequences were determined. These isolates were found to be genotype 1 of lyssavirus and showed the maximum nucleotide sequence homology of 97.6-99.4% with vampire bat-related viruses in Brazil [6]. These results indicate that the Brazilian frugivorous bat rabies viruses in this study are closely related to vampire bat-related viruses that play a main role in rabies virus transmission to livestock in Brazil. KEY WORDS: Brazil, frugivorous bat, rabies.

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“…This genome encodes 5 proteins in the following order from the 3' end: nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA polymerase (L) [15]. The N protein forms an active cytoplasmic ribonucleoprotein (RNP) complex that is essential for viral propagation [20]; there have been many molecular epidemiological studies of N protein [8][9][10][11]. The G protein forms the envelope and capsid (i.e., outer shell) of the virus, which are responsible for viral attachment to the host cell surface and membrane fusion [18].…”

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Genetic and Phylogenetic Analysis of Glycoprotein of Rabies Virus Isolated from Several Species in Brazil

et al. 2004

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ABSTRACT. Genetic and phylogenetic analyses of the region containing the glycoprotein (G) gene, which is related to pathogenicity and antigenicity, and the G-L intergenic region were carried out in 14 Brazilian rabies virus isolates. The isolates were classified as dogrelated rabies virus (DRRV) or vampire bat-related rabies virus (VRRV), by nucleoprotein (N) analysis. The nucleotide and amino acid (AA) homologies of the area containing the G protein gene and G-L intergenic region were generally lower than those of the ectodomain. In both regions, nucleotide and deduced AA homologies were lower among VRRVs than among DRRVs. There were AA differences between DRRV and VRRV at 3 antigenic sites and epitopes (IIa, WB+ and III), suggesting that DRRV and VRRV can be distinguished by differences of antigenicity. In a comparison of phylogenetic trees between the ectodomain and the area containing the G prot ein gene and G-L intergenic region, the branching patterns of the chiropteran and carnivoran rabies virus groups differed, whereas there were clear similarities in patterns within the DRRV and VRRV groups. Additionally, the VRRV isolates were more closely related to chiropteran strains isolated from Latin America than to Brazilian DRRV. These results indicate that Brazilian rabies virus isolates can be classified as DRRV or VRRV by analysis of the G gene and the G-L intergenic region, as well as by N gene analysis.

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Discrimination between dog-related and vampire bat-related rabies viruses in Brazil by strain-specific reverse transcriptase-polymerase chain reaction and restriction fragment length polymorphism analysis

Cited by 49 publications

References 32 publications

Molecular Epidemiological Analysis of Bat Rabies Viruses in Brazil

Molecular Epidemiological Analysis of Bat Rabies Viruses in Brazil

Genetic Characterization of Rabies Viruses Isolated from Frugivorous Bat (Artibeus spp.) in Brazil

Genetic and Phylogenetic Analysis of Glycoprotein of Rabies Virus Isolated from Several Species in Brazil

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