Emerging Pattern of Rabies Deaths and Increased Viral Infectivity

Sharon L. Messenger, Jean S. Smith, Lillian A. Orciari, Pamela A. Yager, Charles E. Rupprecht

Disclosures

Emerging Infectious Diseases. 2003;9(2) 

In This Article

Methods

Sequences were obtained from reverse transcriptase polymerase chain reaction (RT-PCR)-amplified portions of the nucleoprotein (positions 1,157-1,477) regions based on the reference strain M13215.[17] The taxa include 32 rabies virus samples obtained between 1958 and 2000 from frozen (n=27) and formalin-fixed (n=5) U.S. human brain tissue specimens, 17 of 41 U.S. bat species (n=54), and 98 nonhuman terrestrial mammals (24 cats, 3 dogs, 5 cattle, 9 horses, 1 sheep, 1 llama, 42 foxes [grey, red, and kit], 1 raccoon, 1 ringtail cat, and 11 striped skunks), determined to have been infected by a bat rabies virus by using monoclonal antibody screening. Reservoir species from the eight U.S. terrestrial rabies-endemic regions (n=24) were included as outgroups. Nucleotide sequences included in this study will be deposited in GenBank but are also available as an aligned matrix (click here to view).

Phylogenetic analyses used PAUP* 4.0b2,[18] employing the neighbor-joining search algorithm (minimum evolution) using maximum likelihood to estimate Ti:Tv ratio and nucleotide base frequencies (settings correspond to Hasegawa-Kishino-Yano [HKY] 1985 model of nucleotide sequence evolution). We assessed tree support using the nonparametric bootstrap method (1,000 replicates). Phylogenetic analyses presented here included only frozen human brain samples (n=27). Additional analyses (not shown) that included incomplete taxa (formalin-fixed human case samples) did not alter tree topology but decreased nonparametric bootstrap proportions for those nodes including formalin-fixed taxa.

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