We conducted a prospective study in consecutively admitted patients with cirrhosis and ascites in any of the following clinical situations: patients with SBP (Group I), patients admitted for the treatment of uncomplicated ascites without bacterial infections (ASC), as determined by negative microbiological culture and no presence of bacterial DNA in blood and ascitic fluid (AF), who were not receiving long-term SID with norfloxacin as secondary prophylaxis of SBP (Group II) and patients

undergoing SID with norfloxacin as secondary prophylaxis of SBP (Group III). No patient included in the study showed multinodular hepatocellular Lumacaftor mw carcinoma, portal selleckchem thrombosis, alcoholic hepatitis, previous liver transplantation, or previous transjugular intrahepatic portosystemic shunt.

SBP was defined as the presence of >250 polymorphonuclear (PMN) cells/μL in AF. The ethics committee of the hospital approved the study protocol, and all patients gave informed consent to participate in the study. Blood and AF were obtained from all patients at admission and were analyzed for routine biochemical and cytological studies. In patients with SBP, samples were obtained at the time of SBP diagnosis. Blood and

AF cultures were performed in all cases, as described elsewhere.11 Aliquots of blood and AF were inoculated under aseptic conditions in sterile, rubber-sealed Vacutainer SST Org 27569 II tubes (BD Diagnostics, Belgium) that were never exposed to free air. To detect the presence of bacterial DNA fragments in blood and AF, a broad-range polymerase chain reaction (PCR) was performed according to the methodology described elsewhere.12 A quantitative chromogenic limulus amoebocyte lysate test (BioWhittaker, Nottingham, UK) was followed to evaluate endotoxin levels in blood and AF samples as previously described.13 Samples and reagents were handled in an airflow chamber and processed with pyrogen-free material tested by the manufacturers. PMN cells from peripheral blood samples were isolated with PolymorphPrep (Axis-Shield PoC, Oslo, Norway) according to manufacturer’s instructions. After PMN cell isolation, cells were washed twice with freshly made phosphate-buffered saline (PBS) at 4°C. Cell viability was evaluated by trypan blue (Sigma, Madrid, Spain).