Benedict emphasised the Biblical account of creation in his Easter Vigil homily on Saturday, saying it was wrong to think at some point "in some tiny corner of the cosmos there evolved randomly some species of living being capable of reasoning and of trying to find rationality within creation, or to bring rationality into it".
"If man were merely a random product of evolution in some place on the margins of the universe, then his life would make no sense or might even be a chance of nature," he said.
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"But no, reason is there at the beginning: creative, divine reason."
Church teaching holds that Roman Catholicism and evolutionary theory are not necessarily at odds: A Christian can, for example, accept the theory of evolution to help explain developments, but is taught to believe that God, not random chance, is the origin of the world.
The Vatican, however, warns against creationism, or the overly literal interpretation of the Biblical account of creation.
Benedict's voice seemed hoarse and he coughed several times during the lengthy service, which usually ends after midnight.
It was the second late night in a row for the 84-year-old pontiff following his participation in the Good Friday Way of the Cross procession at Rome's Colosseum, which commemorates Jesus' death.
On Sunday, he celebrates Easter Mass in St Peter's Square and gives his traditional Easter greetings in dozens of languages - his last major celebration before next week's beatification of Pope John Paul II.
The Easter Vigil is the most important liturgy on the church's calendar, when the faithful mark the passage from Christ's death to his resurrection on Easter Sunday.
It is rich with symbols, fire and light signifying Jesus' resurrection, and the water used to baptise people into the faith.
On Saturday night, Benedict baptised six adults from Switzerland, Albania, Russia, Peru, Singapore and China, pouring water over their heads as he prayed.
Benedict began the service by lighting a candle and walking down a darkened central aisle of St Peter's Basilica in silence, while hundreds of faithful in the pews shared the flame from candle to candle until the entire basilica twinkled.
