A goal from Andy Lindergaard inside the opening two minutes inflicted Luton's first defeat in eight games against struggling Cheltenham.
It means the Hatters are likely to loose vital ground on the relegation threatened teams in League One after the weekend programme.
This was a fixture that Luton really had to get something from, with Cheltenham just two points and one place above them in the League One table going into the game.
But Luton couldn't wipe out that early goal, as Lindegaard poked home from close range after the Hatters defence didn't deal with Alan Wright's long throw.
However, the Hatters certainly had their chances with Matthew Spring twice denied by excellent saves from Cheltenham goalkeeper Shane Higgs and Marc Wilson also going close in the first half.
Calvin Andrew was then denied with the last touch of the game as his goal bound header was kept out by the impressive Higgs who was at full stretch to his right.
The defeat means Luton are still searching for their first league away win this season, a statistic which will have to change if the Hatters are going to avoid relegation.
Luton's starting line-up had a slightly unfamiliar look to it with three first-team regulars missing.
Hatters boss Kevin Blackwell stated in his pre-match press conference that he had injury concerns, and as a result there was no David Bell and Jaroslaw Fojut in the 16-man squad.
That meant Keith Keane and Wilson both returned to the starting line-up with Paul Furlong also returning from suspension at the expense of Andrew, who dropped to the bench.
Cheltenham gave Guy Madjo his first start for the Club, who replaced Paul Connor, in the only change from their 2-0 defeat at Leyton Orient last week.
Luton's line-up saw Keane move in at centre-back, Matthew Spring push forward as a supporting striker, whilst Dave Edwards and Darren Currie occupied both the flanks.
The Hatters had barely settled into their stride when they were quickly behind to Cheltenham's first attack of the game.
Inside the second minute, former Aston Villa defender Alan Wright launched a long throw into the penalty area which was flicked on by Shane Duff and eventually touched home from close range by Lindegaard, for his first goal of the season.
But Luton's bad start didn't stop them from coming close on a couple of occasions inside the opening ten minutes.
Firstly, on six minutes, Wilson saw a close range header, from Currie's inswinging corner out on the right, tipped neatly over the crossbar by an acrobatic save from Higgs.
The former Bristol Rover shot-stopper was then called upon four minutes later when he was at full stretch to keep out Spring's 25-yard pile driver that was certainly creeping into the far corner.
Chances were continuing to be created at either end and on 17 minutes, Dean Brill kept his side in the game when he got behind a Damian Spencer effort.
The Cheltenham man drilled a low shot from 20-yards that bobbled off the Luton goalkeeper and into the path of Jerry Gill, but the former Birmingham man failed to find enough power in a header after clattering with Madjo.
Luton almost took advantage of Cheltenham's missed opportunity by storming down the other end and going close when Spring's 20-yard right footed curler was splendidly tipped over the crossbar by the outstretched arms of Higgs.
The Hatters were certainly playing the better football, with their clever build-up play deserving more reward as a final product.
Currie then rattled the crossbar on 39 minutes when his corner from the right eluded everyone and almost caught Higgs flatfooted.
AT HALF TIME: CHELTENHAM 1 V 0 LUTON
The second half started in a scrappy manner, but Cheltenham should have doubled their advantage five minutes after the restart.
Luton's defence was split wide open and left Steve Gillespie racing through onto a precise pass. As the striker surged towards the Hatters goal, Brill kept his nerve to make a vital save, which was mopped up by Richard Jackson off the line.
The Hatters also had a chance to get back on level terms two minutes later, but Edwards wasted a golden opportunity by blasting over the crossbar from 12-yards, after Cheltenham failed to deal with Currie's cross into the penalty area.
Blackwell needed to make a change and threw on Drew Talbot on 61 minutes as Luton went back to a 4-4-2 formation.
The visitors were struggling to find their fluency from the first half, as chances were hard to come by.
Spring fizzed a shot towards the Cheltenham goal on 72 minutes, but couldn't force a save from Higgs whilst Talbot almost scored with a header soon afterwards.
Luton were throwing men forward as Andrew was brought on as a third striker. The plan nearly worked when Anthony Grant sent over a cross from the right that was headed inches wide by Talbot on 81 minutes.
Luton then came desperately close to a late equaliser when Higgs spectacularly kept out a Andrew header with a splendid save to his right
AT FULL TIME: CHELTENHAM 1 V 0 LUTON
Cheltenham: Higgs, Gill, Duff, Caines, Gillerspie, Spencer, Bird, Lindergaard, Sinclair, Wright, Madjo (Connor 83)
Subs not used: Brown, Connolly, Gallinagh, D'Agostino
Bookings: Gillespie (43), Sinclair (70), Spencer (80)
Luton: Brill, Perry, Coyne, Currie, Jackson (Andrew 81), Robinson (Talbot 61), Edwards (Grant 66), Keane, Spring, Furlong, Wilson
Subs not used: Goodall, McVeigh
Bookings: Spring (20), Currie (64), Perry (88)
Referee: Mr S J Bratt
Attendance: 3,702 (355 from Luton)
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