Assessment of a quantitative 5 ' nuclease real-time polymerase chain reaction using groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil

作者：Benevenute Jyan Lucas; Dumler John Stephen; Ogrzewalska Maria; Rodrigues Roque Andre Luiz; Califre Mello Victoria Valente; Marques de Sousa Keyla Carstens; Goncalves Luiz Ricardo; D'Andrea Paulo Sergio; de Sampaio Lemos Elba Regina; Machado Rosangela Zacarias; Andre Marcos Rogerio^*

来源：Ticks and Tick-Borne Diseases, 2017, 8(4): 646-656.

DOI：10.1016/j.ttbdis.2017.04.011

New genotypes of Anaplasmataceae agents have been detected in wild carnivores, birds and deer in Brazil. The present work aimed to investigate the presence of Ehrlichia and Anaplasma species in rodents sampled in Brazil. Additionally, a newly designed quantitative 5' nuclease real-time multiplex PCR for Ehrlichia and Anaplasma spp. detection based on groEL gene amplification was designed, showing high specificity and sensitivity (10 groEL fragment copy/mu L). Between 2000 and 2011, different rodent species [n = 60] were trapped in 5 Brazilian biomes. Among 458 rodent spleen samples, 0.4% (2/458) and 2.4% (11/458) were positive for Ehrlichia and Anaplasma spp., respectively. Of 458 samples, 2.0% (9/458) and 1.1% (5/458) were positive for Anaplasma sp. and Ehrlichia sp., respectively, using conventional 16S rRNA PCR assays. Maximum Likelihood phylogenetic analyse based on a small region of 16S rRNA genes positioned the Anaplasma genotypes in rodents near Anaplasma phagocytophilum or Anaplasma marginate and Anaplasma odocoilei isolates. Ehrlichia genotypes were closely related to E. cants. There was a low occurrence of Anaplasma and Ehrlichia in wild and synanthropic rodents in Brazil, suggesting the circulation of new genotypes of these agents in rodents in the studied areas.

• 出版日期2017
• 单位Univ Estadual Paulista