title: Detection and identification of bacterial DNA in patients with cirrhosis and culture-negative, nonneutrocytic ascites.
abstract: The current pathogenic theory of spontaneous bacterial peritonitis (SBP) in patients with cirrhosis and ascites suggests that repeated episodes of bacterial translocation (BT) from intestinal lumen to mesenteric lymph nodes followed by systemic seeding are the key steps for the final development of infectious events. However, most of the episodes of systemic bacterial circulation remain undetected. Therefore, we investigated the hypothetical presence of bacteria in blood and/or ascitic fluid (AF) from patients with cirrhosis and sterile (culture negative) AF by means of bacterial DNA (bactDNA) detection and identification. Twenty-eight consecutively admitted patients with cirrhosis and presence of AF were included in the study. BactDNA was detected using a polymerase chain reaction (PCR)-based method. The corresponding bacteria were identified by nucleotide sequencing of purified PCR products. BactDNA was detected simultaneously in blood and AF in 9 patients (32.1%). DNA sequencing allowed the identification of Escherichia coli (n = 7) and Staphylococcus aureus (n = 2). In all cases, the similarity between the sequence found in AF and blood indicated that the bactDNA present in both locations originated from a single clone (single translocation event). Child-Pugh score and basic hemodynamic, clinical, endoscopic, and biochemical characteristics were similar among patients with or without the presence of bactDNA. In conclusion, we have detected bactDNA in serum and AF in 32% of all patients studied, and this likely represents single clone episodes of translocation and systemic seeding. E. coli is the most frequently identified bacteria.
authors.affiliation: Unidad Hepática, Hospital General Universitario, Universidad Miguel Hernández, Campus de San Juan, San Juan de Alicante, C/Pintor Baeza s/n, 03010 Alicante, Spain. such_jos@gva.es
authors.forename: José
authors.initials: J
authors.lastname: Such
authors.forename: Rubén
authors.initials: R
authors.lastname: Francés
authors.forename: Carlos
authors.initials: C
authors.lastname: Muñoz
authors.forename: Pedro
authors.initials: P
authors.lastname: Zapater
authors.forename: Juan A
authors.initials: JA
authors.lastname: Casellas
authors.forename: Ana
authors.initials: A
authors.lastname: Cifuentes
authors.forename: Francisco
authors.initials: F
authors.lastname: Rodríguez-Valera
authors.forename: Sonia
authors.initials: S
authors.lastname: Pascual
authors.forename: Javier
authors.initials: J
authors.lastname: Sola-Vera
authors.forename: Fernando
authors.initials: F
authors.lastname: Carnicer
authors.forename: Francisco
authors.initials: F
authors.lastname: Uceda
authors.forename: José M
authors.initials: JM
authors.lastname: Palazón
authors.forename: Miguel
authors.initials: M
authors.lastname: Pérez-Mateo
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001201:Ascites / Q000382:microbiology*; D001202:Ascitic Fluid / Q000382:microbiology; D018988:Bacterial Translocation; D004269:DNA, Bacterial / Q000032:analysis* / Q000097:blood; D016099:Endoscopy, Gastrointestinal; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D004932:Esophageal and Gastric Varices / Q000175:diagnosis; D005260:Female; D006801:Humans; D007422:Intestines / Q000382:microbiology; D008103:Liver Cirrhosis / Q000382:microbiology*; D008198:Lymph Nodes / Q000382:microbiology; D008297:Male; D008643:Mesentery; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial