Multiplex PCR for screening of integrons in bacterial lysates

Bacterial integrons are a useful PCR amplification target in epidemiological surveys of bacterial antibiotic resistance, and a variety of primers have been published. We describe multiplex PCR methodology to test for classes 1, 2 and 3 integron-associated integrases in boiled lysates of Gram-negative bacteria. We report on performance in Acinetobacter spp. (n = 50), Enterobacteriaceae (n = 76), Pseudomonas aeruginosa (n = 15), Bacteroidesspp. (n = 69), and in undifferentiated mixed cultures derived from perineal swabs (n = 50) and endotracheal aspirates (n = 8). This method achieved 100% sensitivity and specificity in simple lysates made from a range of bacteria, without requiring DNA extraction, and is recommended as an efficient screening tool for surveys of integron cassettes.