Department of Laboratories, Ospital ng Makati, Makati City, Philippines.
Mem Inst Oswaldo Cruz. 2010 Mar;105(2):117-22. doi: 10.1590/s0074-02762010000200001.
The use of Gram type-specific PCR on buffy coat from clinical specimens for the detection of bacteraemia was evaluated for the first time using whole blood culture as the gold standard. In addition, the established buffy coat culture and whole blood PCR were also compared. Gram-positive bacteria belonging to six species and Gram-negative bacteria from 10 species were isolated and identified by culture and detected using broad-range 16S rDNA primers and Gram-specific primers. Data from the three methods all conferred very high sensitivity, specificity, positive and negative predictive values when compared to whole blood culture. The Kappa coefficients of agreement were 0.9819 (buffy coat PCR), 0.9458 (whole blood PCR) and 1.0 (buffy coat culture), which establishes their validity as alternative methods to routine blood culture in detecting bacteraemia. In addition, results showed that there was a direct correlation of WBC counts greater than 12,000 cells per mm(3) to the occurrence of bacteraemia as detected by the four methods (p < 0.05).
首次使用全血培养作为金标准，评估了在临床标本的白细胞层上使用革兰氏阳性菌和革兰氏阴性菌特异性 PCR 检测菌血症的效果。此外，还比较了建立的白细胞层培养和全血 PCR 方法。通过培养分离并鉴定了属于六种革兰氏阳性菌和十种革兰氏阴性菌的细菌，并使用广谱 16S rDNA 引物和革兰氏特异性引物进行检测。与全血培养相比，这三种方法的数据在检测菌血症时均具有非常高的灵敏度、特异性、阳性预测值和阴性预测值。Kappa 系数的一致性分别为 0.9819（白细胞层 PCR）、0.9458（全血 PCR）和 1.0（白细胞层培养），这证明它们是替代常规血培养检测菌血症的有效方法。此外，结果表明，白细胞计数大于 12000 个细胞/mm(3)与四种方法检测到的菌血症发生直接相关（p < 0.05）。
