<strong>Evaluation of the Time Period for Which Real-Time Polymerase Chain Reaction Detects Dead Bacteria</strong>
Journal Title: Polish Journal of Microbiology - Year 2014, Vol 63, Issue 4
Abstract
Real-time polymerase chain reaction (PCR) is currently used widely for the diagnosis of infectious diseases. We evaluated the time required for which real-time PCR can detect bacteria that had been killed [i]in vitro[/i] by different treatments. Methicillin-resistant [i]Staphylococcus aureus[/i] (MRSA), [i]Staphylococcus epidermidis,[/i] and [i]Escherichia coli[/i] each was treated with alcohol-, antibiotics-, and heat-treatment. Inactivation state of DNA in each sample was confirmed by real-time PCR, including methicillin-resistant [i]Staphylococcus[/i] (MRS)-PCR targeting the [i]mecA[/i] gene and universal PCR targeting 16S rRNA gene. Each strain was confirmed to be culture-negative after treatment. The time for which MRS-PCR could detect viable MRSA treated by alcohol, vancomycin, linezolid, and heat was confirmed to be less than 16, 8, 12, and 8 weeks, respectively. The time period of universal PCR for detection of viable [i]S. epidermidis[/i] treated by alcohol, cefazolin, and heat was less than 4, 20, and 20 weeks, whereas that for viable [i]E. coli[/i] were 8, 20, and 4 weeks, respectively. Our studies reveal that the real-time PCR method can detect DNA in killed bacteria for several weeks, but this capability decreases with time and is lost at 20 weeks after treatment.
Authors and Affiliations
Hyonmin Choe, Yutaka Inaba, Naomi Kobayashi, Yushi Miyamae, Hiroyuki Ike, Hiroshi Fujimaki, Taro Tezuka, Yasuhide Hirata, Tomoyuki Saito
Keywords
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