Ms Rousseff had 55.8% of the votes against 44.2% for centrist candidate Serra with more than 97% of the total counted.
She will be sworn in as President on January 1 next year.
Ms Rousseff, 62, a career civil servant who has never held an elected office before, was bolstered by the support of President Luiz Inacio Lula da Silva.
As polls opened, Ms Rousseff paid tribute to Mr Lula, and assured Brazilians that while he would not have an official role in her government, he would always be near.
"President Lula, obviously, won't be a presence within my cabinet," she said at a final campaign stop in her hometown of Belo Horizonte.
"But I will always talk with the president and I will have a very close and strong relationship with him.
"Nobody in this country will separate me from President Lula."
Ms Rousseff pledged to continue Mr Lula's popular social programmes which have helped take 20 million Brazilians out of poverty since he took office in 2003.
With about 135 million voters, Brazil's powerful economy has helped it achieve an important position as a key emerging nation.
Latin America's largest country is also a world leader in bio-fuel development, while simultaneously working to develop massive deepwater oil fields.
Ms Rousseff will lead a nation due to host the 2014 World Cup and the 2016 Summer Olympics, by which time it is expected to be the globe's fifth-largest economy.
"I want to unite Brazil around a project not just of material development, but also of values," she said.
"When we win an election, we must govern for all Brazilians without exception."
Mr Serra, 68, is a former governor of Sao Paulo state and one-time national health minister who was heavily defeated by Mr Lula in the 2002 presidential election.
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