Pechorsky Alina, Nitzan Yeshayahu, Lazarovitch Tsilia
The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel.
J Microbiol Methods. 2009 Sep;78(3):325-30. doi: 10.1016/j.mimet.2009.07.008. Epub 2009 Jul 16.
Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Acinetobacter baumanii, and Klebsiella pneumoniae were found to be the most prevalent bacteremia-causing bacteria in a survey in a medical center. A PCR method for identification of these five most common pathogens in blood cultures was developed. A unique sequence was chosen for each pathogen and used for primer design. Sixty-one blood samples (from hospitalized patients) in which bacterial growth was detected were processed in parallel by conventional microbiological methods and by the PCR method. The results obtained by PCR were identical to those obtained by conventional methods in 93.4% of the cases. PCR failed to identify bacteria which were found conventionally in only 6.6% of the cases (mostly bacteria not included in the PCR cassette). Another group of eighty-eight blood samples from patients were processed immediately upon their arrival at the laboratory by taking aliquots for the PCR method. The blood sample bottles were processed in parallel by conventional methods. In 78.4% of the cases the results of both methods were identical. In 12.5% of the cases, PCR afforded identification of bacteria but conventional methods showed no bacteria in the sample. On the other hand, PCR afforded 9.1% negative results while conventional methods identified bacteria not included in the PCR cassette. It is concluded that the molecular method appears to be a specific and precise method for identifying pathogenic bacteria in blood samples.
在一家医疗中心的一项调查中发现,金黄色葡萄球菌、表皮葡萄球菌、铜绿假单胞菌、鲍曼不动杆菌和肺炎克雷伯菌是最常见的导致菌血症的细菌。开发了一种用于鉴定血培养中这五种最常见病原体的聚合酶链反应(PCR)方法。为每种病原体选择了一个独特的序列并用于引物设计。对61份(来自住院患者)检测到细菌生长的血样,同时采用传统微生物学方法和PCR方法进行处理。PCR获得的结果在93.4%的病例中与传统方法获得的结果一致。PCR未能鉴定出仅在6.6%的病例中通过传统方法发现的细菌(大多数细菌不在PCR检测盒中)。另一组来自患者的88份血样在到达实验室后立即通过取等分试样进行PCR方法处理。血样瓶同时采用传统方法进行处理。在78.4%的病例中,两种方法的结果一致。在12.5%的病例中,PCR鉴定出了细菌,但传统方法显示样本中没有细菌。另一方面,PCR得出9.1%的阴性结果,而传统方法鉴定出了不在PCR检测盒中的细菌。得出的结论是,分子方法似乎是一种用于鉴定血样中病原菌的特异性和精确性方法。