A bishop had planned to temporarily transfer the saint's relics to another cathedral, around three miles away, but this provoked anger from local parishioners.
Msgr Giuseppe Piemontese had planned to temporarily transfer the saint's relics from the San Valentino Basilica in Terni, Umbria, to another cathedral for Valentine's Day.
But a group of Christians objected, and descended on the basilica.
They seated chairs around a glass coffin containing the remains and holding hands to form a human barrier so that the coffin could not be moved, according to The Daily Telegraph.
Saint Valentine, known as St. Valentine of Rome, served in the third century and was martyred and buried to the north of Rome, his remains were transferred to San Valentino Basilica in 1618.
It was reported that people were angry because they felt their town had been left abandoned in recent years.
One local told the newspaper they "had lost everything" and that removing St. Valentine, a tourist attraction, would not help the area.
Another protestor said: "We are staying here together with our patron saint.
"No-one is going to take him away.
"If they really want to do it, they're going to have to get past us, because we're not moving even one step backwards."
The remains of St. Valentine were due to be moved for a special Valentine's Day celebration in honour of the Catholic Church's Holy Year of Mercy, where people are encouraged to be kind and forgive.
Msgr Giuseppe Piemontese said the remains would have been returned quickly to his birth town and it is thought that he had paid for a special car himself to have them transported for the day of love.
The remains stayed in San Valentino Basilica on February 14th and were not moved.
Msgr Piemontese accused protesting parishioners of behaving in an "intolerant, arrogant and disrespectful manner" at his Sunday service, he also suggested other motives may have been behind the protest.