Adjudication of the final diagnosis One month after study participation, the selleck chemical final diagnosis (SBP) and the aetiology of ascites were independently adjudicated in a blinded fashion by two board-certified gastroenterologists not involved in the patients�� care. Their final assessment was based upon available medical records, including PMN count and the results of all diagnostic investigations, as well as the patient��s response to treatment. Current recommendations were followed[6,7]. The two physicians designated the aetiology of ascites by choosing one or more of the following diagnoses from a standardized list: liver cirrhosis (alcoholic, chronic hepatitis, non-alcoholic steatohepatitis, hemochromatosis, primary biliary cirrhosis, others to be specified), hepatocellular carcinoma, cholangiocellular carcinoma, liver metastasis, peritoneal carcinomatosis, right heart failure, nephrotic syndrome, and others to be specified.
If more than one cause of ascites was identified, the leading disorder responsible for the current episode was established and recorded. Any disagreements in the final diagnosis of a given study participant were resolved by consensus with a third clinician who was considered an expert in the field and recruited to independently review and adjudicate the cases. Paracentesis Paracentesis was performed under aseptic conditions with the patient in the supine position and the puncture site in the left or right lower quadrant. Prior to needle insertion, ultrasound was performed to visualise the intra-abdominal structures.
No study participant suffered complications related to the abdominal puncture procedure. All samples for diagnostic testing were immediately collected at the bedside and processed by laboratory personnel without further delay. Specifically, aliquots of approximately 1mL ascites were centrifuged for 15 min at 500 �� g. The supernatant phase was transferred to a fresh tube and stored at -20 ��C until analysis by ELISA or POC, which occurred within 72 h. Blood samples were also obtained at this time. The ascites samples were used to measure total cell count, PMN count, calprotectin, albumin, total protein, glucose, and lactate dehydrogenase. In addition, two 10 mL aliquots of ascites were subjected to bacterial culture (bottle method) respectively. The serum-ascites albumin gradient (SAAG) was calculated as the difference of albumin in serum and albumin in ascites.
Differential cell count and cytopathology All laboratory analyses were performed in the Central Laboratories BL (Sch?nenbuch, Switzerland) by senior laboratory personnel blinded to patient history and calprotectin levels. Total and differential leukocyte cell counts were determined Batimastat by the manual method using a Neubauer chamber and gentian-violet staining (Leukotic?; bioanalytic GmbH, Freiburg, Germany). The Central Laboratories BL is accredited according to ISO/IEC 17 025 and ISO 15 189 standards.