Blackburn striker Roque Santa Cruz shattered Portsmouth's bid for a top five finish as his second half goal sealed a 1-0 win at Fratton Park on Sunday.
Santa Cruz struck with 16 minutes left to claim the 17th league goal of an impressive first season in English football and leave Portsmouth four points behind fifth placed Everton with two games to play.
A win for Everton in Sunday's late match against Aston Villa would end Portsmouth's hopes of qualifying for the UEFA Cup through their league position.
Harry Redknapp's team will have to ensure European football comes to Fratton Park for the first time by winning next month's FA Cup final against Cardiff.
Blackburn, in eighth place, could still play in Europe next season, although they will probably have to qualify for the UEFA Cup via the Intertoto Cup.
“We saw that we hadn't played good enough to win the game in the first half but we were a different team after half-time,” Santa Cruz said. “It was a nice play for the goal and a great finish.
“We have achieved most of what we wanted this season and it would be nice to qualify for Europe as well. We are still in with a chance.”
David James missed his first league game since joining Portsmouth in 2006 due to the calf injury he suffered last weekend, so Jamie Ashdown made a rare start.
Ashdown looked nervous in the first few minutes, fumbling a cross under pressure from Santa Cruz and Jason Roberts, but Lassana Diarra bailed out his keeper with a hurried clearance.
Portsmouth knew only a win would keep them in the hunt for a top five finish and they started to force the pace.
Sulley Muntari's first-time shot deflected just over off team-mate Nwankwo Kanu. Then Muntari shot wide after Blackburn failed to clear a corner.
Ashdown gradually found his feet and saved well when David Bentley flicked a clever shot towards the top corner.
Portsmouth striker Jermain Defoe had the ball in the net a minute later after running through the centre onto a through pass by Glen Johnson only to be flagged offside.
Defoe threatened again when he found space on the edge of the penalty area and fired a low drive just wide.
Blackburn went straight up the other end and nearly broke the deadlock when Roberts lashed in a powerful effort. Ashdown was equal to it but his parry fell for Santa Cruz, who was unable to get a shot off before being crowded out.
Ashdown had to save Portsmouth again in the 54th minute when New Zealand defender Ryan Nelsen headed Morten Gamst Pedersen's cross goalwards.
Portsmouth had a golden chance when Defoe's cross flashed into Muntari's path but the midfielder couldn't stretch to turn it in.
When Blackburn finally made the breakthrough in the 74th minute it was no surprise that Santa Cruz was the man to deliver the killer blow.
Pedersen's pass was flicked on by Santa Cruz to Roberts, who deftly returned the ball to his strike partner.
Santa Cruz's lethal form left little doubt about the outcome as the Paraguay forward calmly steered his shot past Ashdown.
Diarra forced Brad Friedel to make a fine save with a stoppage time header but Blackburn held on.
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