The visitors clinched the Spartan South Midlands League Division 1 title in midweek but on Saturday the Amps handed them their first defeat since early November to earn three priceless points.
A week after their poor Amersham display, Town returned to the winner's enclosure with a tremendous 1-0 triumph that ensured the club of a play-off place with three games to go.
The new champions were given a guard of honour from their hosts but the pleasantries stopped there and Sam Holmes first half goal proved decisive as Ampthill came out on top in the battle for crucial points.
It was the third time since Christmas the sides had met with Town losing there in the Trophy, but after being the only side to take a point at New Roker, Lee Hawkes masterminded a hugely important win.
Sam Holmes has had a lean time on the goal scoring front of late but he chose a good time to add to his tally when he broke the deadlock and poached the winner in the 22nd minute. Town won a throw-in and Charlie Cullen's set piece move caused problems in the Stotfold defence and it fell for Holmes to hook a volley past Liam Gooch for this 16th goal of the campaign.
Stotfold began the game with plenty of aggression and found the home defence in better form although two low crosses into the goalmouth in a five minute spell around the half hour mark went unpunished.
Ross Tomkins was called on to make some good saves late in the firs half though James Younger headed over and Luke Smith went over from an away free kick.
The Roker men asked plenty of questions after the break but found Town equal to the task and had all the answers. Joe Sutton put a volley wide with 55 minutes gone and later in the half Town keeper Tomkins kept his side ahead with two marvellous stops.
Ampthill had few openings in the second period but with Stotfold going for a leveller the hosts continued to preserve their advantage with a hard working showing and they challenged for every loose ball right until the end to secure a great win over the new champions.
Town will now look to gather momentum in their last three games and look for points to move further up the table to fight for a home tie in the play-offs. They play their last two away games of the season this week making the short trip to Bedford FC on Saturday before going to Letchworth GC Eagles on Tuesday.