The phenomenon of bacteria resistance to three or more antibiotic classes has been defined as multidrug resistance (MDR). Antimicrobial resistance is increasingly becoming a global health problem. The aim of this study was to report MDR Escherichia coli prevalence, resistant and virulence genes from poultry in Jos South LGA of Plateau State. Out of 179 cloacal swabs collected from both local and exotic breeds, 99.44% (178/179) were positives for Escherichia coli. Of 178 samples identified culturally 99.44% (177/178) were confirmed to be positive for Escherichia coli with molecular weight of 401 bp. PCR assay also confirmed the presence of genes associated with resistance to TetA, Sul1, AmpC and GyrA. 81.92% (145/177) of the isolates were resistant to Tetracycline, 80.22% (142/177) to Quinolone, 69.50% (123/177) to Sulphanomide and 66.10% (117/177) to Ampicillin. 18 isolates that showed resistance for up to four different antibiotics were further analysed by Multiplex PCR to detect Eae, HlyA, RfbE, FliC and Stx2 virulence genes. 44.4% (15/18) of the isolates were positive for Eae gene, 27.7% (5/18) for Stx2, 22.20% (4/18) for Rfbe gene, 5.5% (1) for Hlya gene, and none for FliC gene. Our results showed high antibiotic resistance, virulent genes and significant levels of MDR in Escherichia coli from poultry. This study underscores the need for antimicrobial stewardship in poultry because of its implication for food safety and health. This is important because poultry farmers form a significant group of small holder farmers in Nigeria.