It was the first time Carlo Ancelotti's side had failed to win in La Liga since the last time the two sides met back in October.
Unai Emery's Sevilla come into the game on a great run of form themselves. Per WhoScored, the last time his side were defeated in the league was at the beginning of February when, coincidentally, Barca came to town.
They will be buoyed further by their qualification for the next round of the UEFA Europa League, however, Madrid's record in Andalusia is a formidable one, with six goals being scored by Los Blancos in two out of their last three visits, per FussballDatenbank.
Seven were put past Emery's side in the reverse fixture earlier this season, so clearly the home side need to be on their game from the first whistle to avoid another hammering.
Date: Wednesday March 26
Time: 9 p.m. GMT/5 p.m. ET
TV Info: Sky Sports 4 (UK) / beIN SPORTS 1 (US)
If there is anything Emery must take from that game, it is to keep things tight and not try to play overly expansive football against a side that can hit you where it hurts to devastating effect.
Karim Benzema's two goals at the weekend weren't enough to win his side the match but on another day he might well have had four or five.
That's only seven less than Sevilla have scored in all competitions.
You can be sure that all three will be fired up for this match, knowing the points are required to keep pace at the top of the league.
But Emery's resurgent side could well throw a spanner in the works, however, and Real Madrid TV's Rhiannon Jones noted Ancelotti's comments:
The win against Osasuna in the last game saw the manager equal Juande Ramos' record of five straight La Liga wins, per WhoScored, and 14 straight games in which they have scored at home is also another club record.
Those records will count for nothing if Los Palanganas cannot stem the tide from Madrid's front three.
Defensive midfield duo Stephane Mbia and Vicente Iborra need to ensure that the spaces in between the lines are kept to a minimum, and they will have to put in an almighty 90-minute shift to see off the danger throughout.
Iborra is particularly combative in the heat of battle but with the threat of goals from set pieces a very real possibility, perhaps he needs to rein in his effusiveness in the tackle just a little.
Unlike Javier Mascherano at the weekend, central defender Federico Fazio is adept at dealing with the high ball into the box, therefore it may favour the away side if they can get Ronaldo and Bale into the wider areas to play the ball in behind.
Sevilla's hopes of springing a surprise will fall to their effervescent captain Ivan Rakitic. The talisman has weighed in with 11 goals and nine assists in his 26 appearances (one as sub), per WhoScored.
It's a fine return from a central midfield position and from a player that continues to lead by example.
A second loss in as many games for Madrid might well see the top three change positions yet again, in one of the most exciting title races La Liga has seen in many years.