The test was carried out on the players after their 1-0 defeat against African champions Algeria in Friday’s build-up game.
The result revealed all the Super Eagles players in camp are free from the virus and are available for selection.
“Ahead of tonight’s friendly game against Tunisia, all Super Eagles players have tested negative to Covid-19,” read a tweet from the NFF.
Nigeria will hope to continue their dominance over the Carthage Eagles in the encounter, having not lost to the side since 2004.
The Nigeria Football Federation (NFF) had previously debunked rumours making the rounds that some of the Super Eagles players tested positive for the virus before their clash against Algeria.
“Please discountenance any reports that some of our players tested positive to Covid-19 ahead of tomorrow’s friendly game against Algeria. It is a blatant falsehood,” NFF wrote in a statement.
“The writer of the story, who was not part of the press conference earlier today, totally misrepresented what coach [Gernot] Rohr said that since the outbreak of the pandemic, four of our players had tested positive but were all now fully recovered from the virus and back in action.”
Nigeria have lined up the friendlies as part of their preparations for the Africa Cup of Nations qualifying game against Sierra Leone in November.
The Super Eagles lead the Group L table with six points from two games after victories over the Republic of Benin and Lesotho.