Judges, including rock star Steven Tyler from Aerosmith, picked the winner from a total of 86 contestants at the show, watched by millions of viewers around the world.
Last year's winner Olivia Culpo of the United States placed a diamond crown on the head of Isler, who wore a silver rhine-stone studded evening dress.
Isler's triumph was the seventh Miss Universe win for Venezuela, where beauty pageants are a source of national pride.
After the show, she beamed and waved to journalists, wearing the Miss Universe sash and standing next to the show's owner, US tycoon Donald Trump.
"I have a lot of emotions. I can't describe all the things that I feel at this moment because I'm shaking," she said.
"I'm still in shock. I feel so blessed, so happy to be here."
All 86 contestants initially appeared on stage in red and black cocktail dresses. They then changed into red and black bikinis.
The judges then whittled down the contestants to a final 16, who faced off in a bikini round, carrying white fur stoles.
Then a final 10 women strutted on the stage in elaborate sequinned evening gowns, before the final five answered questions in the interview round.
Host country Russia was eliminated immediately, prompting boos in the audience.
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