Multiplex PCR for the detection of tetracycline resistant genes

Specific primer pairs were selected for the PCR amplification of 14 tetracy cline resistant genes commonly found in Gram positive and Cram negative org anisms. Combinations of primer pairs were used in multiplex PCR reactions t o detect specific groups of tel genes as follows; Group I: tet(B), tet(C), tet(D); Group II: tet(A), tet(E), tet(G); Group III. tet(K), tet(L), tet(M) , tet(O), tet(S); Group IV: tetA(P), tet(Q), tet(X). To test: the multiplex PCR, Groups I and II were used on 25 clinical isolates of Salmonella enter ica serovar Typhimurium DT104. Group III primers were used to investigate 1 9 clinical isolates of methicillin-resistant Staphylococcus aureus. Multipl ex PCR should result in significant savings in terms of labour and cost in analysis of a large number of strains when compared with using an individua l PCR for targeting each gene. It may also be a useful method to differenti ate the types of tetracycline resistance when used as an additional marker for the purpose of outbreak investigation and surveillance.