25º Congresso Brasileiro de Microbiologia

Marcus Vinicius Aragão Batista (UFPE); Tiago Alessandro Espinola Ferreira (UFRPE); Antonio Carlos de Freitas (UFPE); Valdir de Queiroz Balbino (UFPE)

Influenza A virus subtype H1N1 is the most common cause of flu in humans, and some strains are endemic and others are seasonal. There are some endemic strains in pigs (swine Influenza) and in birds (avian Influenza). Nowadays, a novel Influenza A virus subtype H1N1 acquired new ability to spread between humans efficiently, and it became pandemic. Thus, it is important to develop more rapid and accurate detection tests. However, as the number of genetic information increases, some modifications in the methodology should improve the efficiency of the tests. So, this study aims at developing a new approach for design primers and probes for the detection and characterization of Influenza A virus, mainly the subtype H1N1. This work consisted on the analysis of Influenza A sequences deposited in the database of the National Center for Biotechnology Information (NCBI) in the last three years. The sequences of all genes were aligned using an incorporated version of the program ClustalW in the Molecular Evolutionary Genetics Analysis Software version 4.0 (MEGA4). The entropy, which shows the complexity of the variation among the sequences, was calculated for each site, and the regions with the higher values were used for the primers and probes design. One set of primers was designed for universal detection of type A Influenza virus, one for specifically detect all swine type A Influenza virus, and one for specifically detect swine H1N1 Influenza virus.The employed methodology allowed the identification of regions that presented good levels of conservation, apparently sufficient for the attainment of degenerate and also type-specific primers and probes that are already being validated experimentally. Primers and probes produced in this work presented criteria of specificity and sensitivity superior to those described in the literature, resulting in the increment of the capacity of identification of the viral types circulating in Brazil.

Financial support: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).