Following an injury scare that required medical intervention that was picked up while playing with Betis, Guardado was selected in the final 23-man squad for the 2018 World Cup, his first as captain. In the first match, he played 74 minutes in a 1–0 defeat of Germany, going on to take part in all the fixtures including the round of 16 loss with Brazil. Following their elimination, it was reported by Mexican sports newspaper "Récord" that he had played through the tournament with a muscle injury.