'Do the Conga' was a top 10 hit in 1984. The lyrics have been reworked to give information on discounts available to those wanting to save money on train travel.
Ads will feature a cross-section of commuters clad in gold Lycra dancing to the Black Lace classic.
Black Lace released 'Agadoo' in 1984. It became a gigantic hit and a favourite of wedding receptions the world over.
Iain Hildreth, director of marketing at thetrainline.com said passengers can get the best deals if they buy tickets before they travel. He said: "People tell us that they don't always remember to do this so our new campaign is designed to help remind them in a colourful, catchy and instantly memorable way.
"With Black Lace on vocals and that infamous dance favourite 'The Conga', we're confident our new campaign will certainly get that message across and encourage more people to take advantage of the savings on thetrainline.com to get out and about in the UK by train."
Creative is by DLKW Lowe and media by Vizeum, and replaces the "sheep ads", which first aired two and a half years ago.
The first ad will break on 30 April during Britain's Got Talent on ITV1 followed by 10 second spots, supported by radio and online campaigns including homepage takeovers of MSN, Sky and YouTube where a special 'Choo Choob' channel has been created, featuring behind the scenes footage, outtakes and further versions of 'Do the Conga'.
Black Lace tracks have been used in ads before – 'Agadoo' was used to encourage recycling in 2000.
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