By Centers for Disease Control and Prevention
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Extra info for Emerging Infectious Diseases - Vol. 14, No. 6, June 2008
Sample text
Tissues were fixed in 10% neutral buffered formalin and embedded in paraffin; 5-μm sections were stained with hematoxylin and eosin for histopathologic diagnosis. Unfixed tissues for virologic and molecular analyses were stored at –80oC. , Danbury, CT, USA). , Valencia, CA, USA) according to manufacturer’s instructions and eluted in a final volume of 60 μL of buffer. Total RNA was also extracted from lung tissue collected from specific-pathogen–free dogs without respiratory disease. Real-Time Reverse Transcription–PCR A single-step quantitative real-time reverse transcription–PCR (RT-PCR) was performed on total RNA extracted from the canine tissue samples by using the QuantiTect Probe RT-PCR Kit containing ROX as a passive reference dye (QIAGEN).
The canine viruses differentiated them from contemporary equine viruses. The substitution of lysine for asparagine at position 54 maintains the positive charge of the residue and is of unknown functional significance. Position 83 located within antigenic site E of human H3 has been implicated in antigenic drift (11). The substitution of serine for asparagine at position 83 in canine H3 maintains the polar nature of the residue, but the functional significance in the evolution of canine influenza is not immediately apparent.
Gov/eid • Vol. 14, No. 6, June 2008 907 RESEARCH from humoral immune responses (11,15,16). Although no evidence supports high variability at positions 118 and 261, neighboring positions 121 (antigenic site D) and 262 (antigenic site E) are sites of frequent positive selection in human H3 genes (11). The 2005 isolates from nongreyhound dogs differed from each other by substitution of proline for serine at position 107 in the HA1 subunit of A/canine/ Jacksonville/2005. Serine is conserved at position 107 in all other equine and canine isolates except for A/equine/ Jilin/1/89, which has a threonine substitution (17).