Strootman had already received minor damage to his right knee during the Netherlands' 2-0 friendly defeat to France last Wednesday and was substituted at half-time as a precautionary measure.
However, the 24-year-old had recovered in time for the weekend's Serie A clash—only to then tear cartilage in his left knee before being stretchered off just 13 minutes into the Stadio San Paolo encounter.
Whilst the injury is a blow to his Italian club side, the impact on his national side is far greater. Rudi Garcia's Roma will still secure Champions League qualification without Strootman but for Louis van Gaal's Netherlands the situation causes a huge dilemma.
As per BBC Sport, van Gaal had named the midfielder as one of three players—alongside Bayern's Arjen Robben and Manchester United's Robin van Persie—guaranteed a seat on the plane to Brazil and his absence will leave a huge hole in Holland's World Cup plans.
Van Gaal conceded: "For the national side it is a huge blow. Kevin made a huge contribution to our qualification, he never let us down and now his dreams have died."
The former PSV midfielder was superb in his country's unbeaten World Cup qualification campaign, completing more passes than any other Dutch player and also scoring his third international goal against Hungary. Although he is an extremely able technician, perhaps it is his physical presence that will be missed most.
Strootman has previously been labelled as the "Dutch Roy Keane" and is one of the game's true warriors. With the likes of Wesley Sneijder and Robben in the side, it is important to have a real fighter to sure things up in the middle of the park; the Roma man was easily van Gaal's best option prior to Sunday's injury.
The hole may be plugged by the experienced Nigel de Jong but the 29-year-old Milan player is not quite to the standard of Strootman.
Four years ago, de Jong helped Holland to the World Cup final in South Africa and even played an instrumental part in keeping Spain at bay until extra-time before falling to that Andres Iniesta strike late on.
However, in South Africa he was sat next to the experienced Marc van Bommel so he could share the defensive responsibilities. In Brazil, that might not be the case as van Gaal will more than likely opt for a slightly more attacking approach than his predecessor Bert van Marwijk. De Jong isn't the player he was at Manchester City and would unquestionably struggle against the might of Spain.
Against their other Group B opponents (Chile and Australia), Strootman's absence should not cause too many problems. The main issue lies with the potential last 16 match. Should the Dutch finish behind Spain then it is likely that they'll meet Brazil in Belo Horizonte on June 28—that could well be where the Netherlands make their World Cup exit.
The Netherlands eliminated Brazil in South Africa at the quarter-final stage and once again De Jong was a key figure. He was partnered with van Bommel, though, and the hosts are also a lot better than they were four years ago.
Everybody knows that van Gaal's side are very strong going forward and only Germany could better their 34 goals in qualifying. However, despite only conceding five times on the road to Brazil, there are some uncertainties around the back four and their need for a strong holding midfielder cannot be underestimated.
Strootman might not be the most exciting name in world football but he'll certainly be a huge miss for the Dutch. With less than 100 days until the big kick off, manager van Gaal has a genuine dilemma on his hands.
Who should van Gaal pick to fill the hole vacated by Strootman? Post your views below.
Stats from WhoScored.com