Estefan addressed the amped crowd and expressed her gratitude to the gay community.
"I want to thank you so much, because the gay community has been behind me 100%," she said, as seen in the video below. "'Conga' broke through in all the gay clubs before anybody else noticed it was a hot song. You guys have been close to heart and always will be."
She recalled that for her 1994 single "Everlasting Love," she was nine months pregnant and didn't want to be on camera.
She told the Score crowd, "So who could do me better than me? You!"
The crowd responded with shouts of "Las dragas!," referring to the female impersonators Estefan cast for the music video.
"Hotel Nacional" is off Little Miss Havana, Estefan's first English-language album since 2003. Pharrell Williams, who also lives in Miami, produced the album along with Estefan's husband Emilio.
Click below for videos of Estefan's Score appearance as well her music videos.