AmaKhosi, fielding a weakened side, lost 2-1 at ‘home’ in Dobsonville to Ivorian side ASEC Abidjan, who will be heavy favourites for the return leg this coming weekend.
Goals from Zagbayou Eurand and Boua Koffi Davy – either side of Matthew Rusike's strike – means Chiefs need to score twice in Cote d’Ivoire to advance.
And with coach Stuart Baxter likely to send a weakened team again in a competition he has made it clear he has little time for, Chiefs appear to be heading out.
Champions League holders Al Ahly claimed a narrow 1-0 home win over Difaâ Êl Jadida from Morocco that leaves them with a tricky away leg and also possibly heading out.
Midfielder Abdullah Al-Saied netted the only goal of the game for the Egyptians.
Angolan side Petro Atletico have given themselves a chance to advance when they suffered a 2-1 loss at Cameroon outfit Coton Sport, but now just need a single goal at home to reach the group stages.
Real Bamako won the battle of the Malien sides when they defeated compatriots Djoliba 2-1 at home and now face a difficult away journey, though at least it will be to familiar territory.
Zambian champions Nkana were held 0-0 at home by Tunisian side CA Bizertin, who will be favourites for the return leg, while the match between Guinea side Horoya and Etoile du Sahel from Tunisia finished with the same score line with the return to come in North Africa.
AC Leopards from Congo have some breathing space after a 2-0 home win over Medeama of Ghana, while Cote d’Ivoire club Séwé Sport beat Bayelsa Utd from Nigeria at home by the same score line.
The return legs will be played this coming weekend.
CAF CONFEDERATION CUP PLAY-OFF FIRST-LEG ROUND RESULTS:
Nkana (Zambia) 0-0 CA Bizertin (Tunisia)
Al Ahly (Egypt) 1-0 Difaâ Êl Jadida (Morocco)
AS Real (Mali) 2-1 Djoliba (Mali)
AC Leopards (Congo) 2-0 Medeama (Ghana)
Kaizer Chiefs (South Africa) 1-2 ASEC Mimosas (Cote d'Ivoire)
Coton Sport (Cameroon) 2-1 Petro Atletico (Angola)
Horoya (Guinea) 0-0 Etoile du Sahel (Tunisia)
Séwé Sport (Cote d'Ivoire) 2–0 Bayelsa Utd (Nigeria)