The Inter Miami forward has begun the 2024 season in scintillating form, netting five goals and providing two assists in five competitive matches.
However, he was forced to miss Saturday's victory over DC United after picking up an injury in last Wednesday's 3-1 win against Nashville in the CONCACAF Champions Cup.
Messi grabbed a goal and an assist before he was withdrawn in the early stages of the second period due to a hamstring problem.
Speaking after Saturday's clash with DC United, Inter Miami boss Tata Martino revealed Messi's injury will have to be assessed on a "week by week" basis.
Martino told reporters: "Messi's injury must be dealt with week by week, and we will evaluate it. What is clear is that there is a goal with him, which is to play in the CONCACAF quarterfinals. He has a muscle injury. The selection issue is for the people of the national team to talk about."
The World Cup holders have now confirmed that Messi will not be in the squad for their upcoming friendlies due to a "minor" injury issue.
A statement on Argentina's official X page read: "The captain of Argentina, Lionel Messi, will not be able to be in the squad for the friendlies in USA due to a minor injury to the hamstring of his right leg, suffered in his team's Inter Miami CF match against Nashville SC."
As a result, Argentina's all-time leading goalscorer and appearance-maker will miss out on the chance to add to the 106 goals he has scored in 180 international matches.
Despite being unavailable for the current international period, Messi is still expected to have a key role to play in Argentina's defence of their Copa America title, with the competition set to get underway on June 20.
In addition to Messi's absence, Argentina boss Lionel Scaloni has also seen Marcos Senesi, Paulo Dybala and Exequiel Palacios withdraw from the squad due to injury.
Following those withdrawals, Scaloni has opted to add Real Betis midfielder Guido Rodriguez to his squad for the upcoming games in the USA.
Argentina will face El Salvador in Philadelphia on March 22, before they travel to Los Angeles for a meeting with Costa Rica on March 26.