Chipstead hung on to beat Berkhamsted 2-1 on Saturday and knock them out of the FA Trophy at the first qualifying round stage.
With keeper James Weatherill injured in The Comrades’ midweek win at Didcot (see later), Berko boss Steve Bateman made one change for this away trip, bringing in James Shaw for his debut after signing from Hayes & Yeading United.
Max Farrelly was first to have a fierce shot for Berko but it rose over the bar and Matt Bateman hit a shot wide.
Chipstead took the lead in the 28th minute after Manny Folarin lost the ball and conceded a foul just outside the Berko box. Sam Clayton punished it with an expertly flighted free-kick over the wall and into the top corner of the net.
Oliver Paine in the home goal then pulled off a double save to deny Ashley Morrissey and Bateman.
In the second-half Berko were expecting another Clayton shot to go over the bar, but it struck the woodwork and Anthony Oaks was first to react to make it 2-0.
Berko introduced fresh legs in Adiel Mannion and Louis Austin and played better as they tried to rescue the tie. In the 74th minute, a high ball from skipper Dan Murphy went to the back post where Ryan Kinnane challenged and the ball broke to Bateman, who scored for the ninth consecutive game.
That set up a grandstand finish with Murphy driving a shot just over and Liam McCrohan having a chance to equalise and set-up replay, but he placed his shot wide.
Net man Shaw went up for a late corner but to no avail.
It was Berko’s first defeat since September 11, when they went out of the FA Cup to Coggeshall Town.
The Comrades win in midweek to move into league’s upper echelons
Berko moved into the playoff spots of the Evo-Stik South Central Division One table with a fine 4-3 away win at Didcot on Tuesday night.
They made a dream start with an early goal when the referee played a good advantage when Bateman was fouled flicking the ball on and Will Summerfield beat a defender before lobbing the ball over the head of Leigh Bedwell in the home goal.
Midway through the half, Didcot equalised when skipper Adam Learoyd headed home from a Morgan Williams corner.
Morrissey then tricked his way to the byline and was brought down for a penalty, which Bateman converted.
Holding at a dead ball led to a penalty at the other end, but Didcot’s Williams hit the outside of the post to waste it.
Five minutes before the interval Berko took a crucial 3-1 lead. Summerfield was in his own half when he got in front of the centre backs and latched onto a long ball to run clear. Bedwell saved his shot, but the loose ball was rammed home by Bateman.
Ten minutes into the second-half Berko were undone by another set-piece. This time it was to the near post and the ball took two deflections, the last being off James Towell for an own goal.
It didn’t take long for The Comrades to restore their two-goal edge when a Morrissey free-kick from was headed in by Ryan Kinnane.
An ankle injury to Weatherill meant skipper Dan Murphy had to take goal kicks for Berko.
Didcot were awarded another penalty on 80 minutes and this time Williams converted to make it 4-3. That left Berko with a nervous end but they held on for the win.
After a four-match away run, Berko return home to Broadwater this Saturday to host 15th-placed Coleshill Town in the league. They are then at home again on Tuesday night against Aylesbury United in the Southern League Challenge Cup.