Chilwell pulled up in stoppage time of Chelsea’s 2-1 Champions League win over Dinamo Zagreb on Wednesday.
The 25-year-old grabbed the back of his left thigh after making a sprint, falling to the ground before being helped from the field by Chelsea’s medical staff, and was pictured leaving Stamford Bridge on crutches.
“Results show that Ben has suffered a significant injury and the defender is unfortunately expected to miss the World Cup,” Chelsea said in a statement.
“Ben will now begin a rehabilitation programme with the club’s medical department.”
In a statement on social media, Chilwell said: “Following my ACL injury I worked so hard to make sure I was ready for the World Cup. It has always been a dream of mine.
“Unfortunately, this won’t be possible following my scan results. I’ll do all I can to get back playing for Chelsea ASAP.”
Chilwell, who was a part of England’s Euro 2020 squad, missed a chunk of last season due to a ruptured cruciate ligament but had returned to Chelsea’s first team fold this season with 13 appearances in all competitions.
The news comes as another blow to England with the team also set to miss the services of Chelsea’s Reece James, who was ruled out for eight weeks due to a knee injury he picked up in their game against AC Milan.
England are also sweating on the fitness of Manchester City duo Kyle Walker and Kalvin Phillips.
England kick off their World Cup campaign against Iran on Nov. 21.