Kwesi Appiah's side headed into the game one point ahead of their Group D rivals in the race for the only qualification spot into the next stage, but knew that defeat would end their prospects of reaching next year's finals in Brazil.
Marseille attacker Andre Ayew was named in the Ghana starting XI, the 23-year-old featuring for the national team for the first time in six months after taking temporary leave from international football.
However, the hosts had to cope without attacking midfielder Kevin-Prince Boateng, who was unable due to injury, while Southampton striker Emmanuel Mayuka - on loan at Sochaux - started up front for Zambia.
Zambia had yet to lose a game in qualifying going into the contest at Baba Yara Stadium, Kumasi, but it was Ghana who were in control for the most part.
Goals from Majeed Waris and Kwadwo Asamoah gave them the advantage before Emmanuel Mbola pulled one back late on.
The hosts broke the deadlock with their first real chance of the game after 17 minutes, Waris heading in at the near post after Rennes defender John Boye had shown great determination to chase a seemingly lost cause and lift the ball back into the Spartak Moscow striker's path.
Ghana gained confidence after going ahead, and Appiah's men twice went close to doubling their advantage, Waris heading wide before Asamoah Gyan forced Kennedy Mweene into a fine save with a fierce effort from long range.
Zambia failed to create anything of real note in a below-par first-half performance, and they were fortunate not to fall further behind when Wakaso Mubarak sent a free-kick inches wide of the post from the edge of the area.
Herve Renard's side - who won the Africa Cup of Nations in 2012 - looked to try and haul their way back into the game in the early stages of the second half, substitute Fwayo Tembo drawing a save from home goalkeeper Fatau Dauda.
However, the visitors' hopes of qualification seemed to have been extinguished 16 minutes into the second period as Asamoah found the bottom corner with a vicious effort from distance that left Mweene with no chance.
Zambia refused to lie down, though, and gave themselves half a chance 13 minutes later as Emmanuel Mbola rifled home from the edge of the box to fray the nerves of the home fans.
Renard's men pushed tirelessly to attempt to find a leveller and keep their hopes of progression alive, but their efforts proved to no avail as Ghana held on to clinch their spot in the final round, where they will meet one of the other qualifiers in a two-leg play-off for a place in the finals.
