A 1-1 draw against Bolton on Tuesday represented another body blow, although, mathematically, the top six is still within reach.
Here’s what is required for Danny Cowley’s side and the rest of the contenders to reach the League One play-offs this season...
The Blues are the biggest outsiders for a top six finish sitting 10th in League One, 12 points away from sixth-placed Wycombe.
Despite run of one defeat in 11 games, their hopes have since faded after failing to win any of their last four games.
Cowley’s men have at least one game in hand on the teams above them, which could see his side reduce the gap.
However, they require four wins just to reach sixth at this moment in time – even a total of seven straight wins are unlikely to be enough.
Kieran McKenna’s side sit one place above Pompey in ninth, four points clear of their rivals, although have played two games more.
The Tractor Boys fell to their first defeat in 12 on Saturday, which has since slowed their progress up the table.
Town sit eight points away from the play-offs – on 64 points – and realistically must win all their remaining five matches in order to reach the top six.
The U’s have sat comfortably in the top six throughout the campaign, but disappointing losses against Plymouth and Morecambe have seen Karl Robinson’s men drop out away.
United sit eighth in League One, just three points away from the play-offs, and are still contention, but will have to return to winning ways if they want a chance of achieving success.
However, it will be a tough end to the season for the U’s, who face three sides currently above them in the table.
The Black Cats have enjoyed a major resurgence under Alex Neil since his January appointment and are unbeaten in their previous seven league games.
Sunderland sit seventh in the table, just two points behind Wycombe on 70 points with a game in hand, which could lift them as high as fifth.
They face Oxford on Saturday, and must also take on Plymouth, while and Rotherham in their penultimate fixture of the season.
Gareth Ainsworth’s men currently occupy the final play-off position in sixth place, following a run of five wins from seven games.
With five matches to go, the Chairboys must face fourth-placed Plymouth and fellow play-off candidates Sheffield Wednesday.
With a number of teams chasing hard, Wanderers will have to avoid any slip-ups which could potentially hamper their claim play-off qualification.
After flirting outside the top six for most of the season, a run of eight wins from nine games have seen them climb to fifth in the table, also with a game in hand on a number of teams below.
Darren Moore’s side have hit form at the perfect time, but face tough tests against Bolton and MK Dons in their next two outings.
Should the Owls win their game in hand, they could extend the gap between seventh place by as much as seven points, which would all but seal a top-six placing.
Steven Schumacher’s side are the most comfortably sitting in the play-off chase at the moment.
The Pilgrims are fourth in League One, eight points ahead of seventh-placed Sunderland.
However, four of their remaining five games are against teams also involved in the hunt for promotion, with fixtures against Wycombe, Sunderland, MK Dons and Wigan.
Should they drop points against such, it could open the door to the chasing pack, but a win against Burton on Saturday would all but secure their place in the top six.
Rotherham, MK Dons and Wigan have all but confirmed their places in the play-offs, thereby haven’t been included as they continue to scrap it out for two automatic places.