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.
Dillon
B
B
Centre for Infectious Diseases and Microbiology, University of Sydney, Australia.
Thomas
L
L
Mohmand
G
G
Zelynski
A
A
Iredell
J
J
D000150:Acinetobacter / Q000235:genetics* / Q000302:isolation & purification; D001439:Bacteroides / Q000235:genetics* / Q000302:isolation & purification; D004755:Enterobacteriaceae / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D019426:Integrases / Q000737:chemistry / Q000235:genetics; D032023:Integrons / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D019426:Integrases