Real Potosi came from behind on Thursday to win 3-1 at Oriente Petrolero with goals from Leonel Morales, Maximiliano Andrada and Cristian Diaz clinching victory for the visitors, who moved up to third in the standings.
But it was Universitario's 4-1 triumph over San Jose that was the story of round 20, as Javier Vega's victorious side claimed top spot in the table ahead of The Strongest, who were held to a 0-0 draw at Wilstermann.
The Strongest had only moved into first position over the weekend after Universitario were stunned 5-0 by bottom club Blooming.
Universitario started Wednesday's clash in third, behind second-placed San Jose on goal difference, while The Strongest had a one-point lead at the top of the table.
It took just eight minutes for Universitario to open the scoring in front of their home fans in Sucre as Alejandro Bejarano's persistence paid off.
The 29-year-old midfielder chased a back pass to San Jose's goalkeeper, won possession and then converted but the visitors equalised two minutes later thanks to Carlos Neumann's sharp finish.
Bolivia international Gustavo Pinedo then took control of the match for Universitario, tapping home a rebound in the 17th minute and finishing a one-on-one opportunity nine minutes before half-time to put the hosts 3-1 ahead.
San Jose never recovered from Pinedo's blitz and in the 64th minute Lucas Ojeda produced an impressive long-range chip to wrap up the three points for Universitario.
Vega's men moved to 38 points, one ahead of The Strongest, while Real Potosi (36 points) leapfrogged San Jose (35) into third.
San Jose have lost three games in a row.
In other LFPB results, Nacional Potosi trumped Blooming 3-0 and Guabira played out a 1-1 draw with Aurora.
