Evaluation of the time period for which real-time polymerase chain reaction detects dead bacteria.
Evaluation of the time period for which real-time polymerase chain reaction detects dead bacteria.
复制标题
评估实时聚合酶链式反应检测死细菌的时间段。
DOI：
10.33073/pjm-2014-053
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发表时间：
2014
影响因子：
2.1
通讯作者：
Tomoyuki Saito
中科院分区：
文献类型：
--
作者：
H. Choe;Y. Inaba;N. Kobayashi;Y. Miyamae;H. Ike;H. Fujimaki;T. Tezuka;Y. Hirata;Tomoyuki Saito
实时聚合酶链反应（PCR）目前广泛用于感染的诊断。我们评估了治疗后的时间，在此期间，实时PCR可以检测死细菌。通过耐甲氧西林葡萄球菌（MRS）-PCR和通用PCR，通过实时PCR鉴定细菌DNA的存在。耐甲氧西林金黄色葡萄球菌（MRSA）、表皮葡萄球菌和大肠杆菌分别在体外用酒精、抗生素或热处理杀死。经酒精、万古霉素、利奈唑胺和加热处理的MRSA的MRS-PCR检测期分别小于16、8、12和8周。通用PCR的检测周期为10 ~ 20天。酒精、头孢唑啉和热处理的E. epidermidis的存活时间分别小于20、20和4周，而E. coli分别为8周、20周和4周。在成功治疗之前和之后，还通过MRS和通用PCR评估了感染的关节置换术患者中可检测到的细菌DNA的存在。6例患者治疗前MRS-PCR阳性，治疗后平均20.8周（95%可信区间，13.2 - 33.7）后全部转为阴性。通用PCR在治疗后平均15.2周（95%CI，12.4至18.0）时在4例患者中检测到残留细菌DNA，在治疗后平均17.3周（95%CI，10.6至24.0）时在7例患者中检测到阴性。我们的研究表明，实时PCR检测死亡细菌数周，但这种能力随着时间的推移而下降，并可能在治疗后20周失去。
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.