The International Rugby Board has confirmed that the North v South clash of the rugby hemispheres will take place at Twickenham on 30 November.
The Millennium Stadium bid for the game when the fixture was initially put out to tender, but the IRB have now opted to go to London for the inaugural fixture.
"The match was put out to tender, and the WRU duly submitted a bid. It was never at any stage formally agreed that it would be in Cardiff, and we're obviously disappointed to have missed out on this occasion," said WRU media manager Lyn Davies.
South Africa's Andre Watson has already been named as the referee for the game and the IRB will shortly announce a match sponsor and TV broadcaster.
Set to become a biennial event in the rugby calendar, the proceeds will go towards the development of rugby in the smaller playing countries around the globe.
IRB chief executive Mike Miller claims there is "tremendous enthusiasm" for the game.
The IRB will also hold their Annual Awards ceremony in London on the same day as the game.
The WRU has teamed up with Crawshay's to tour Georgia in June. The side, which will be called the Crawshays WRU President's XV, will play Emerging Georgia twice, in a tour which supports the Welsh Government's efforts to grow relations between the two countries.
Wales Sevens are looking to re-group after a disappointing tournament last time out in South Africa. The next double leg in New Zealand and Las Vegas offers Gareth Williams a chance to redeem themselves.
Wales Women's head coach Rhys Edwards was at Llandovery to see the conclusion of an entertaining Women's Regional championship where any one of three teams were in contention for title glory on the last day of action.