The Wildcats fell to defeat for the first time in the FA WSL Spring Series after conceding late on at Doncaster Rovers Belles.
Having fallen behind to Christie Murray’s first-half strike, the Wildcats levelled after the break as Becky Salicki headed home.
But their hopes of taking a deserved point were dashed when Courtney Sweetman-Kirk nodded home a corner to seal victory for the hosts.
Lee Sanders made one enforced change to the side that came from behind to beat Oxford a fortnight ago as Lauren Briggs replaced the injured Nat Gutteridge.
And Durham found themselves on the back foot early on, with Salicki having to be alert to clear off the line after Rachael Laws’ punched clearance fell at the feet of Emily Simpkins.
Laws then had to be at her best moments later to deny Sweetman-Kirk from close range after Murray had pulled the ball back to her.
But the stopper was eventually beaten after 20 minutes, with Murray controlling a cross before firing home from a tight angle.
The Wildcats could have bounced back almost instantly in bizarre circumstances, as Nicola Davies had to keep out an attempted backpass from Kasia Lipka.
Beth Hepple then had a sight of goal after taking the ball past three defenders but saw her effort deflected over the bar, before Durham penalty appeals were waved away on the stroke of half-time.
A deserved equaliser did come just after the hour mark as a training ground routine allowed Salicki to nod home at the back post via a deflection off Leandra Little.
And the Wildcats could have taken the lead moments later – Emily Roberts turning Little before seeing her effort tipped over the bar.
Chances were at a premium during the final stages, with both sides looking to get forward and seal victory.
It was the hosts, however, who snatched the win when Murray’s fine delivery was headed home by Sweetman-Kirk at the far post.
There was no time for the Wildcats to find a leveller and they will now refocus their efforts ahead of the visit of Brighton & Hove Albion next weekend.