The teams head into this one off the back of contrasting results in their most recent outings but the clash promises to be an enthralling one nonetheless.
Match preview
South Sudan fell to a narrow 1-0 defeat against the Leopards of Congo when the sides went head to head at Benjamin Mkapa National Stadium on March 27.
Stefano Cusin's men were on course to come away with a hard-fought point in Dar es Salaam before Gabriel Charpentier netted in the final minute of normal time to settle matters.
As a result, the Bright Stars have dropped to fourth place in Group G, three points behind Congo in second and a further three adrift of leaders Mali.
Next up is the challenge of an opposing outfit who have come out on top in each of the previous two editions of this fixture by a combined score of 3-1.
Having lost each of their last three home matches, Wednesday's hosts will be looking to breathe life into their AFCON qualification journey with a crucial victory here.
Gambia showed great resolve to see off the threat of Mali and claim a hard-fought 1-0 victory in the sides' clash at the Stade Mohamed V on March 28.
Tom Saintfiet's men withstood a lot of pressure in the opening hour of the contest after which they snatched a priceless winner through Omar Colley 11 minutes from time.
With six points amassed from four matches so far, The Scorpions have moved up to third in their qualifying group, level on points with Congo in second.
Currently without a win in any of their last five away games in which they have only managed to score once, Wednesday's visitors now prepare for what is set to be another tough assignment on the road.
South Sudan Africa Cup of Nations Qualifying form:
L
L
W
L
Gambia Africa Cup of Nations Qualifying form:
W
L
L
W
Gambia form (all competitions):
D
W
L
D
L
W
Team News
Peter Manyang missed South Sudan's last outing through suspension but the 29-year-old is now eligible to feature once again and should earn an immediate recall to the XI.
This should see Samuel Taban drop out of the starting lineup after the defender was handed only his sixth international appearance against Congo.
Omar Colley had a game to remember last time out as he opened his Gambia account at the 44th time of asking which also saw him become his nation's most capped player of all time.
Ablie Jallow came up with the decisive moment the last time Wednesday's opponents met and he will be looking to deliver the goods once again.
South Sudan possible starting lineup:
Mawith; Wani, Manyang, Toha, Dhata; Malish, Achol, Chol; Akio, Riak, Kuach
Gambia possible starting lineup:
Gaye; Colley, Sanneh, Sonko; Janko, Manneh, Ngum, Touray; Barrow, Ceesay, Jallow