Evaluation of the Time Period for which Real-Time Polymerase Chain Reaction Detects Dead Bacteria

Abstract

Real-time polymerase chain reaction (PCR) is currently widely used for the diagnosis of infections. We evaluated the time after treatment during which real-time PCR can detect dead bacteria. The presence of bacterial DNA was identified by real-time PCR through methicillin-resistant Staphylococcus (MRS)-PCR and universal PCR. Methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, and Escherichia coli were each killed with alcohol, antibiotics, or heat treatment in vitro. The detection periods of MRS-PCR for MRSA treated by alcohol, vancomycin, linezolid, and heat were found to be less than 16, 8, 12, and 8 weeks, respectively. The detection period of universal PCR for S. epidermidis treated by alcohol, cefazolin, and heat was less than 20, 20, and 4 weeks, whereas that for E. coli was 8, 20, and 4 weeks, respectively. The presence of detectable bacterial DNA in infected arthroplasty patients before and after successful treatment was also assessed by MRS- and universal PCR. MRS-PCR was positive in 6 patients before treatment and all became negative after a mean interval of 20.8 weeks (95% confidential interval, 13.2 to 33.7) after treatment. Universal PCR detected remnant bacterial DNA in 4 patients at a mean of 15.2 weeks (95% CI, 12.4 to 18.0) after treatment and was negative in 7 patients at a mean of 17.3 weeks (95% CI, 10.6 to 24.0) after treatment. Our studies revealed that real-time PCR detects dead bacteria for several weeks, but this capability decreases with time and is likely lost by 20 weeks after treatment.