Tunisian club Esperance failed to create a record unbeaten run in the CAF Champions League after losing 1-0 at ASO Chlef of Algeria.
Cameroon-born striker Anicet Eyenga scored midway through the opening half in Chlef to end a 19-match unbeaten streak by the defending champions, spanning two seasons.
Esperance needed to avoid defeat in Algeria to climb above six-time Champions League title-holders Al Ahly of Egypt, who went 19 matches unbeaten between the 2004 and 2006 seasons.
The Tunisians did overtake Ahly when they defeated compatriots Etoile Sahel last month, but the result was cancelled after the Sousse club were disqualified for pitch invasions.
After losing at ASPAC of Benin last April, the 'Blood and Gold' went 12 games unbeaten to lift the trophy a second time and travelled to Chlef without a loss in seven qualifying and pool games this season.
Apart from the chance of creating a record, there was little at stake, with Esperance and Sunshine Stars of Nigeria already through to the semi-finals from Group A and Chlef doomed to finish last after three consecutive defeats.
Esperance coach Nabil Maaloul seemed most concerned about avoiding injuries and yellow cards as he rested several regulars, including goalkeeper Moez Ben Cherifia, midfielder Youssef Msakni and Cameroonian striker Yannick Ndjeng.
Ahly and four-time champions TP Mazembe from Congo DR have qualified for the semi-finals from Group B and play on Sunday to determine who finishes first and avoids Esperance.
Mazembe lead Ahly on head-to-head results and can clinch first place with a victory at Berekum Chelsea of Ghana, while Ahly face century-old Cairo rivals Zamalek in a simultaneous kick-off.