Komen N, Morsink M C, Beiboer S, Miggelbrink A, Willemsen P, van der Harst E, Lange J F, van Leeuwen W B
Department of Surgery, Erasmus Medical Center Rotterdam, Rotterdam, The Netherlands.
J Microbiol Methods. 2009 Oct;79(1):67-70. doi: 10.1016/j.mimet.2009.08.004. Epub 2009 Aug 14.
A semi-quantitative Real-Time PCR strategy was developed to identify potential indicator organisms for anastomotic leakage in peritoneal drainage fluid, Escherichia coli and Enterococcus faecalis. The analytical performance of the amplification method was validated with 10 culture-positive and 7 culture-negative peritoneal drain fluid samples, obtained from 9 different patients with a colorectal anastomosis. Real-Time PCR results were fully concordant with the microbiological culture results. However, among the culture-negative samples, four false-positive RT-PCR results were found. All false-positives originated from a single patient with a surgical site infection. This may indicate an elevated sensitivity of the RT-PCR method. The results showed that the semi-quantitative RT-PCR method has a clear potential to be useful as a powerful tool in early detection of anastomotic leakage.
开发了一种半定量实时聚合酶链反应（Real-Time PCR）策略，以鉴定腹膜引流液中吻合口漏的潜在指示微生物，即大肠杆菌和粪肠球菌。该扩增方法的分析性能通过从9例结直肠吻合术患者中获得的10份培养阳性和7份培养阴性的腹膜引流液样本进行了验证。实时PCR结果与微生物培养结果完全一致。然而，在培养阴性样本中，发现了4例假阳性RT-PCR结果。所有假阳性均来自一名手术部位感染患者。这可能表明RT-PCR方法的敏感性较高。结果表明，半定量RT-PCR方法具有作为早期检测吻合口漏的有力工具的明显潜力。
