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The Highlanders lost 44-16 to the Waratahs in Sydney and failed to take a competition point from the match.
But results at the weekend mean only a high-scoring close game between the Brumbies and the Force in Canberra this Friday could deny the Highlanders their first finals action since 2002.
The Highlanders are fourth on the table, the same position they were in at the start of the penultimate round.
They could have sealed a playoff spot with any sort of competition point last night, but that never looked likely.
The Highlanders are on 42 points, a point ahead of the fifth-placed Hurricanes, who have the bye next week and cannot get any more points.
The Force, Chiefs and the Brumbies are all on 40 points, meaning they are all still in with a shout.
The Chiefs play the Blues and will get the chance to continue their title defence if they beat the home side at Eden Park on Friday night.
The Force takes on the Brumbies and both could qualify if they get two points each from the game.
If teams finish tied on points, teams with the most wins go through to the playoffs.
The Highlanders fall down in this regard as they have just eight wins, and the Force and the Brumbies have each having bagged one extra victory.
The Highlanders will have the advantage of knowing exactly what they have to do when they play the Crusaders in Christchurch next Saturday night.
If the men from the South do beat the Crusaders, with a bonus point, deny the home team a bonus point, and the Sharks lose heavily to the Stormers, the Highlanders could actually finish second and will have a bye in the first week of the playoffs.
The Waratahs, who have never won the competition, have confirmed top place on the ladder with their win over the Highlanders last night.