Footage of the moment at a Chick-fil-A branch in the US state on North Carolina has since been viewed 86,000 times on social media.
It shows an unnamed manager announcing: "We have one of our team members that is starting surgery right now for breast cancer and we'd like to say a short prayer for her, if you'd like to join us.
"It's Ms Trish, so if you all know her from our dining room, today is a rough day for us, so we're hoping that things go well.
"If you bow your heads and, I'm not great at this, but I'm going to do my best.
'Dear Lord, please just protect Ms Trish as she begins her surgery today.
"Please give the surgeon a steady hand as they perform the needed surgery on her."
The film pans around the chicken restaurant in Belmont, showing families sat at tables in silence with their eyes closed last Friday.
The manager continued: "Please let everybody in the surgery room be at their best today and have sharp minds and clear heads as they work on her.
"We need her back here.
"She is our light, she is the pillar in the community for us and everybody here loves her so much.
"Please, please just protect her and get her back here to brighten our days as soon as you can."
Commenting on the video, one Facebook user called Marcie Fritz said: "It's awesome to see a company stop in the middle of busy lunchtime and say a prayer for their employee!"
Another Facebook user, Carolyn Cockerham, said: "Wonderful to see all these folks bowing in prayer to pray for this special lady."
The founder of Chick-fil-A, S Truett Cathy, who died in 2014, was known as a devote Southern Baptist.
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