Standing next to French President Emmanuel Macron and his wife Brigitte, the US president could be seen poking his left forefinger at the back of the first lady’s hand as his wife remained unresponsive.
Mr Trump tried to wrap his finger around his wife’s but she again appeared to snub his hand-holding efforts.
He remained unperturbed and made a more concerted effort to clutch her hand before she surrendered to his advances.
The first couple, who welcomed Mr Macron and his wife to the White House for the Trump administration’s first state visit this week, have had other embarrassing hand-holding encounters.
In May, Ms Trump was filmed clearly swatting away her husband’s hand during a trip to Tel Aviv, Israel. In February, the billionaire property developer once again attempted but failed to hold hands with his wife as the pair left the White House for a trip to Ohio.
The latest hand holding saga has been mocked on social media and late night TV show host Stephen Colbert poked fun at the incident.
“Trump tried to hold Melania’s hand and she is not having it man,” The Late Show host said. “He’s like trying to arouse a dead trout.”
“You can’t blame her though. I mean, she watches 60 Minutes. She knows where that hand has been.”
The Daily Show dubbed the moment with music from Steven Spielberg thriller Jaws - a film about a giant man-eating great white shark attacking beachgoers.
“Look at poor Melania right hand trying to tell Trump that he didn’t dig deep enough,” quipped one Twitter user.
“Poor Melania, she looked like she was holding back tears under her hat as she held Trump’s hand,” added another.
Mr Macron’s three-day visit to Washington has also seen Mr Trump brush off the French leader’s “dandruff”.
"They're all saying what a great relationship we have, and they're actually correct," Mr Trump said to Mr Macron and gathered press in the Oval Office during a meeting between the pair.
"We do have a very special relationship. In fact, I'll get that little piece of dandruff off - we have to make him perfect. He is perfect."
Mr Trump then turned to Mr Macron and brushed his shoulder.
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