The 31-year-old American's stunning vault in Albuquerque, New Mexico, eclipsed Isinbayeva's indoor mark of 5.01m set in 2012 and is sure to heat up their rivalry ahead of August's outdoor world championships in Moscow.
"I was in a groove tonight," Suhr, who set the mark on her first attempt at the height, said.
"I knew some big things could happen, I just need to be healthy and come down the runway.
"I'm glad that I got 5 metres out of the way, (with the) 5.02, because it is a mental barrier."
Suhr, who denied Isinbayeva a hat-trick of Olympic golds at the London Games, made three unsuccessful attempts at 5.07m to surpass the Russian's outdoor world record of 5.06m.
A former basketball player, Suhr had been jousting for years to supplant the seemingly invincible Isinbayeva, training in an under-heated shed during the frigid winters of her native upstate New York.
She was runner-up when Isinbayeva set a world record to win gold at the 2008 Beijing Olympics, but turned the tables in London as the Russian struggled to a disappointing bronze.
"I knew if I could execute and stay focused and listen to the instructions I had that it could happen," Suhr said of her record jump.
"I guess I knew where I was in my training, I just wasn't going to let everyone else know."
Prior to Suhr's 5.02m vault, which secured her ninth national record in six years, Isinbayeva held eight indoor jumps superior to the American's personal indoor best of 4.88m set last year.
Daylight separates Suhr's outdoor best from Isinbayeva's, the Russian having surpassed the American's 4.92m 10 times.
But Americans are on the rise, Suhr said.
"The US is starting to bring up the women's vault and I think you can see it," she said. "Around the world they are going to see the results."
Until her unsuccessful bid for 5.07m, Suhr had not missed a single attempt on Saturday, clearing her first height at 4.65m, and again at 4.70m and 4.75m.
She soared over the bar at 4.80m and upped the ante to 4.90m, smashing her previous indoor best before her record-breaking vault.
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