Harriers, who went into the match well clear of the relegation zone following an undefeated four match run, were beaten by a second half goal from Dan Maguire which lifted his side to the top of the table.
The defeat means the hosts have now failed to win on home soil since triumphing 3-1 against Hereford on December 26.
They have also been unable to keep a clean sheet at Aggborough since October 22 when they overcame Farsley Celtic 2-0 on October 22.
Kidderminster boss Russ Penn, a former York player, made one change from the side which fought out a goalless draw at Gateshead the previous Saturday.
Defender Alex Penny, whose previous clubs included a brief spell at Stourbridge, was drafted in for his debut after moving to Aggborough from Boston United.
The twenty three year old’s appearance meant Ryan Johnson was dropped to the substitutes’ bench.
Harriers, who drew 1-1 at York in early November, went close in the ninth minute when a curling free kick by former York campaigner Ashley Hemmings swerved just wide of the far post.
New boy Penny was then involved in a promising build up which resulted in Alex Prosser firing wide from some 20 yards out.
York responded with a good move of their own when Maguire was neatly played through, but he was unable to get in a meaningful shot while being put under pressure from Sam Austin.
Maguire was a threat again a couple of minutes later but this time he was foiled by quick thinking Harriers’ goalkeeper Will Mannion, who came out to save at the striker’s feet.
Harriers were soon back up field and a cross by Ollie Shenton was kicked out by Paddy McLaughlin for a corner which was taken by Prosser and headed inches over the bar by Milan Butterfield.
With minutes Austin had an effort well saved by visiting goalkeeper Pete Jameson before the loose ball reached Penny whose fierce goalbound shot was well blocked by David Ferguson.
Butterfield then had a good chance to score just after the half hour mark but he wasted the opportunity by heading Keith Lowe’s lofted cross into the ground and wide of the target.
The keenly contested first half drew to a close with Chambers shooting high over the bar from distance.
The Minstermen grabbed what proved to be the winner a minute into the second period when the unfortunate Penny attempted to head back to Mannion, but the ball fell well short and Maguire nipped in to rifle home.
It was a sickening blow for Harriers who quickly set about trying to get back on terms, but were regularly frustrated by the well organised York rear-guard.
The visitors, however, went close to extending their slender lead when substitute Jordan Burrow was not far off the mark with a close range header.
Harriers responded with a glancing header by Chambers, but the ball flashed well wide before Butterfield had a pile-driver blocked in front of goal.
The midfielder then had another effort which dipped just over the bar before Hemmings had a powerful shot expertly tipped over the bar by Jameson in the dying minutes.
Kidderminster: Mannion; Penny, Moyo, R. Williams (Johnson, 65), Lowe, Austin; Butterfield, Prosser Shenton (E. Williams, 61); Hemmings, Chambers (Samuels, 80). Subs not used: Palmer, Freemantle.
York: Jameson; Bond, Tait, Newton, McNulty, Ferguson; Moke, McLaughlin (Griffiths, 40), Green; Maguire (Burrow, 62), Willoughby (Kempster, 79). Subs not used: King, York.
Referee: Richard Holmes.
Attendance: 1,574 (281 away fans).