Sablikova turned in a blistering time of four minutes 2.53 seconds to win by more than two seconds and become the second Czech woman to win a Winter Games gold following cross-country skier Katerina Neumannova in 2006.
Stephanie Beckert of Germany took the silver medal in 4:04.62. Canadian Kristina Groves won the bronze with 4:04.84.
"I felt good going into the race," an elated Sablikova said. "It was a difficult race. There were three pairs after me and I hoped to get at least one medal. I have one medal now and everything is possible (for the 5000)."
The 22-year-old Sablikova, long and lean with a smooth, gliding style, made up for her Olympics disappointment at the Turin Games, where she finished seventh in the 3000 and fourth in the 5000m.
Sablikova could hardly contain her joy, fist-pumping with her coach in the infield when it became clear victory was hers and then jumping with excitement with her arms straight up when she reached the top of the podium.
The Czech, rail-thin by comparison to her more muscular rivals at 5-foot-7 and 119 pounds, set a daunting target time in the 11th of 14 pairs and watched as her chief rivals faltered.
First Groves, roared on by the Canadian faithful urging her on in hopes of seeing the host country's first gold of the Games, and then 2006 champion Ireen Wust of the Netherlands pushed hard and put themselves ahead of Sablikova's pace.
However, both faded in the last laps, Groves hanging on to snare the bronze and still win thunderous cheers from the crowd.
Wurst finished seventh in 4:08.09, 5.56 seconds back.
Place of birth: Nove Mesto Na Morave, Czech Republic
Residence: Velky Osek, Czech Republic
Previous Olympic results:
Fourth place, 2006 Winter Olympics, 5000m
Seventh place, 2006 Winter Olympics, 3000m
Gold medallist, 2009 World Allround Championships, small combination
2x gold medallist, 2010, 2007 European Championships, small combination
2x bronze medallist, 2009, 2008 European Championships, small combination
Sablikova began skating in 1998 and has also been competing in road cycling since 2007. Her younger brother, Milan, is also a speed skater.
A lack of artifical ice tracks in her homeland has forced her to train on short tracks or travel abroad. In 2007 she won the national speed skating title competing on a frozen lake.
Her Vancouver victory made her just the second Czech woman to win a Winter Games gold following cross-country skier Katerina Neumannova in 2006.