Department of Gastroenterology, Hospital de la Santa Creu i Sant Pau, Universitat Autònoma de Barcelona, Institut d'Investigacions Biomèdiques Sant Pau, Spain.
Aliment Pharmacol Ther. 2011 Jan;33(2):275-84. doi: 10.1111/j.1365-2036.2010.04506.x. Epub 2010 Nov 11.
Despite inoculation into blood culture bottles, ascitic fluid culture is negative in 50% of cases of spontaneous bacterial peritonitis (SBP).
To determine whether 16S rDNA gene detection by real-time polymerase chain reaction (PCR) and sequencing increases the efficacy of culture in microbiological diagnosis of spontaneous bacterial peritonitis.
We prospectively included 55 consecutive spontaneous bacterial peritonitis episodes in cirrhotic patients, 20 cirrhotic patients with sterile ascites and 27 patients with neoplasic ascites. Ascitic fluid was inoculated into blood culture bottles at the bedside and tested for bacterial DNA by real-time PCR and sequencing of 16S rDNA gene.
Bacterial DNA was detected in 23/25 (92%) culture-positive SBP, 16/30 (53%) culture-negative SBP (P = 0.002 with respect to culture-positive SBP), 12/20 (60%) sterile ascites (P = 0.01 with respect to culture-positive SBP) and 0/27 neoplasic ascites (P < 0.001 with respect to other groups). Sequencing identified to genus or species level 12 culture-positive SBP, six culture-negative SBP and six sterile ascites. In the remaining cases with positive PCR, sequencing did not yield a definitive bacterial identification.
Bacterial DNA was not detected in almost half the culture-negative spontaneous bacterial peritonitis episodes. Methodology used in the present study did not always allow identification of amplified bacterial DNA.
尽管在血培养瓶中进行接种,但在 50%的自发性细菌性腹膜炎(SBP)病例中,腹水培养呈阴性。
确定实时聚合酶链反应(PCR)和测序的 16S rDNA 基因检测是否提高了自发性细菌性腹膜炎微生物诊断中培养的效果。
我们前瞻性地纳入了 55 例连续的肝硬化患者自发性细菌性腹膜炎发作,20 例肝硬化无菌性腹水患者和 27 例肿瘤性腹水患者。将腹水在床边接种到血培养瓶中,并通过实时 PCR 和 16S rDNA 基因测序检测细菌 DNA。
在 25 例培养阳性的 SBP 中,有 23 例(92%)检测到细菌 DNA,在 30 例培养阴性的 SBP 中,有 16 例(53%)检测到(与培养阳性的 SBP 相比,P=0.002),在 20 例无菌性腹水中有 12 例(60%)(与培养阳性的 SBP 相比,P=0.01),在 27 例肿瘤性腹水中没有检测到(与其他组相比,P<0.001)。测序鉴定出 12 例培养阳性的 SBP、6 例培养阴性的 SBP 和 6 例无菌性腹水为细菌属或种水平。在其余 PCR 阳性的病例中,测序未得出明确的细菌鉴定结果。
在近一半的培养阴性自发性细菌性腹膜炎病例中未检测到细菌 DNA。本研究中使用的方法并不总是能够鉴定出扩增的细菌 DNA。