Searches from the UK dropped by 24 per cent, while global searches decreased by nine per cent.
There was also a four per cent decrease in searches for all future hotel stays in London - suggesting the decrease is both caused by and limited to the Tube Strike, the company said.
Trivago compared searches made for hotel stays during the industrial action this week with corresponding days during the previous week.
“It is extremely rare to see a significant decrease in travel interest to a city as large as London,” commented Denise Bartlett, from Trivago UK Public Relations.
“With around 3000 hotels bookable via trivago, a decrease of 24 per cent is likely to have a significant effect on the city’s tourism over the two-day period.
“The decrease in hotel searches is just one indicator of the effects of the tube strike, with visitors relying almost solely on the underground system in order to travel within the city,” she said.
The industrial action over pay and rosters for the new night tube this week was the largest strike by London Underground workers for 13 years.
Hospitality businesses have welcomed the introduction of the 24 hour service, which is predicted to boost late night business.
London Mayor Boris Johnson told the BBC yesterday that he did not 'give a stuff' when the night service came in to effect, but that it would definitely happen despite the dispute with London Underground workers.
Unions have warned that further action could follow this summer unless London Underground improves its offer to employees.