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Harlequins' 10-match winning streak came to an end as London Wasps held on for a 17-16 victory at the Twickenham Stoop.
Nick Evans kicked a late penalty to give Quins a 42-40 success when the same sides met on the opening day of the season but failed to repeat the trick, this time his miss in the 78th minute sealed their first defeat in all competitions since November.
The result also sees Quins fall one point behind Leicester Tigers at the top of the Aviva Premiership and moves Wasps up to fourth.
Wasps raced into a 14-0 lead with two tries in the first 13 minutes, one each for wingers Tom Varndell and Christian Wade.
Varndell intercepted an Evans' pass to go in under the posts and Wade finished off a passing move for his 17th league try of the season, a score that was confirmed by the TMO. Stand-off Nicky Robinson, who had missed an early penalty shot, converted on both occasions.
Evans, Robinson's opposite number, had also been off-target with his first penalty attempt but corrected that error to his side on the board in the 18th minute after Wasps inside centre Chris Bell was sent to the sin-bin for a high tackle on Nick Easter.
Quins took advantage of their extra man to get back in the contest, inside centre Matt Hooper gathering a loose ball for the score after Wasps full-back Hugo Southwell failed to clear.
Evans added the conversion and brought the first half to a close with his second penalty to reduce Quins' deficit to 14-13 at the break.
Wasps produced a resolute defensive effort in the second half and Robinson's 60th-minute penalty padded their advantage to four points.
Evans set up an exciting finish with his third penalty on 72 minutes, taking his personal haul to 11 points, but he failed with a final chance late on as Wasps survived.
|Friday 18th April|
|Harlequins 24 - 20 Leicester|
|Friday 11th April|
|Sale Sharks 12 - 27 Harlequins|
|Friday 4th April|
|Amlin Challenge Cup|
|Stade Francais 6 - 29 Harlequins|
|Saturday 29th March|
|Harlequins 23 - 9 London Irish|
|Saturday 22nd March|
|Saracens 39 - 17 Harlequins|
|Saturday 1st March|
|Harlequins 21 - 20 Worcester|
Read the thoughts and opinions of Stuart Barnes with skysports.com