Vos, the Olympic road race champion, put in a decisive attack on the final ascent of the legendary Cauberg climb to win her first world title since taking the gold medal as an amateur back in 2006.
Australia's Rachel Neylan took silver after crossing the line 10 seconds behind Vos, while Italian Elisa Longo Borghini took bronze a further eight seconds back.
The top three were part of a seven-rider breakaway that formed when both Vos and Borghini jumped clear of the peloton on the Cauberg during the sixth of eight 16-kilometre loops to join a break of five riders that had formed off the front in the previous lap.
Having finished runner-up in the previous five successive world championships, Vos was in no mood to fall short once again - especially with the whole of the Netherlands craving victory.
Described by the great Eddy Merckx as "the finest cyclist of [her] generation", Vos could call on the assistance of 19-year-old team-mate Anna van der Breggen who, riding her debut world championships, had forced her way into the initial break.
The peloton - led by Britain's Emma Pooley - tried to react after Vos's first attack 34km from the finish. But soon the gap had ballooned to two minutes for the seven escapees and it was game over for the likes of Britain, Belgium, Russia and France, all of whom had no riders in the mix.
Like Vos, Borghini had an Italian team-mate in the shape of rookie Rossella Ratto. Besides Neylan and van der Breggen, the break also included Charlotte Becker of Germany and American Amber Neben, who had made the initial attack with 70km remaining.
After a subdued start to the 129km race, things had got spicy in the Province of Limburg when a huge pile up on the second lap saw numerous riders hit the deck and held up around two thirds of the peloton.
Amongst those caught up in the melee were the 2009 world champion Tatiana Guderzo of Italy, and Germany's Judith Arndt, the 2004 world champion, who was riding her last race as a professional days after retaining her time trial world title.
Briton Katie Colcough crashed heavily and cut her face. She was later forced to retire alongside a whole host of riders.
But the early crash was a distant memory when the leaders rode onto the Cauberg two hours later for the penultimate ascent at the end of lap seven.
Vos put in a huge attack on the Alpe d'Huez of Holland, with only Borghini and Neylan able to follow. Neben and van den Breggen managed to claw their way back to the leaders, but Ratto and Becker were both out of the equation.
Borghini led the quintet over the line as the bell sounded for the final lap, almost three minutes ahead of the resigned peloton.
Vos and van der Breggen controlled the pace on the front up the Bemelerberg climb until Borghini signalled for a race car and dropped back. Vos, seemingly curious, slowed to shadow her main rival, while van der Breggen momentarily rode off the front.
But it was all back together when, with 3km remaining, the five leaders approached the Cauberg for the final time.
Neylan put in the first attack to shed both van der Breggen and Neben, before Vos herself powered away with the race's decisive move.
Borghini exploded, leaving only Neylan to chase the red-hot favourite. But things kept to the script as Vos extended her lead over the top of the summit in front of the ballistic home fans.
Vos had time to take a Dutch flag from the crowd before crossing the line to cap off an extraordinary year with a much-belated second world championships title - and, astonishingly, her first as a professional.
"It is so special to race in your own country," said Vos, before paying tribute to her team-mate van der Breggen, who "did a really good job to keep us in front".
"I knew it could be the decisive break and it was great to see the gap growing from one, to two and then to three minutes," said Vos. "When we got to the last lap I thought this is the moment when I could be world champion."
The 25-year-old from North Brabant said she could now take stock of a triumphant season which has also seen her win Olympic gold, the world and Dutch national cyclo-cross championships and the women's Giro d'Italia.
"In the last few weeks I've been focussing on this and not thinking too much about the Olympic gold. Now I can start to look back and reflect on what I have done," she said.
Neylan and Borghini completed the podium, with Neber crossing the line 33 seconds down in fourth and van den Breggen cheered home for fifth at 55 seconds.
The peloton, including fifteenth-place Pooley, crossed the line four minutes and 47 seconds off the pace.