The Israeli metropolis won 43% of the votes, leaving New York City behind in the second place with only 14% of the votes.
The top 10 cities also included Toronto, Sao Paulo, Madrid, London, New Orleans and Mexico City. Voting was held last month on the company's website.
The competition included additional categories, including Best Nightlife (won by New York City), Best Pride (San Francisco) and Best Sand & Sun (Sydney).
Contest's website. Middle East's gay capital
Tel Aviv, which was marked as a favorite even before the voting concluded, was described during the competition as "the gay capital of the Middle East, exotic and welcoming, with a Mediterranean c'est la vie attitude."
Shai Doitsh, brand manager of the Tel Aviv Gay Vibe tourism campaign, told Ynet shortly after learning of the results: "This makes us very proud.
"This is the peak of six years of activity and further proof that the decision made by the Tourism Ministry and Tel Aviv Municipality to invest in gay tourism and put their faith in the program we built was the right decision.
"This is an excellent start for the coming year of activity, in which we will continue to innovate and surprise, and mainly bring thousands of tourists to Israel," he added.