OBJECTIVETo evaluate the value of ascitic bacterial 16S rRNA gene determination in the rapid diagnosis of spontaneous bacterial peritonitis (SBP).

METHODS16S rRNA gene from bacterial DNA in ascites was determined by quantitative fluorescent polymerase chain reaction (PCR) in 76 patients with suspected SBP and 6 patients with non-infectious ascites. The results were compared with those obtained from bacterial culture.

RESULTSThe positive rate of SBP was 22.4% among patients detected with ascitic bacterial 16S rRNA gene determination-based quantitative fluorescent PCR, which was significantly higher than that (7.9%) in patients only received bacterial culture (P<0.05). In addition,in 6 patients with non-infectious ascites,both the 16S rRNA gene determination-based quantitative fluorescent PCR and bacterial culture showed negative results.

CONCLUSIONS16S rRNA gene determination-based quantitative fluorescent PCR can be an effective tool for the rapid diagnosis of SBP. It is more sensitive than the bacterial culture.

Adult ; Aged ; Ascitic Fluid ; microbiology ; Bacterial Infections ; diagnosis ; DNA, Bacterial ; analysis ; Female ; Humans ; Male ; Middle Aged ; Peritonitis ; diagnosis ; microbiology ; RNA, Ribosomal, 16S

OBJECTIVETo evaluate the significance of determining ascitic bacterial 16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP).

METHODSAscitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases. The results were compared with ascitic bacterial culture simultaneously.

RESULTSOf 76 ascitic samples, 17 were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative(G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P < 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of non-infectious ascites with chronic liver diseases.

CONCLUSIONSDetermination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.

Adult ; Aged ; Ascitic Fluid ; microbiology ; Bacterial Infections ; diagnosis ; microbiology ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; microbiology ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Peritonitis ; diagnosis ; microbiology ; Polymerase Chain Reaction ; methods ; RNA, Bacterial ; isolation & purification ; RNA, Ribosomal, 16S ; isolation & purification

Spontaneous bacterial peritonitis (SBP) is a common complication of cirrhosis. Based on our current understanding of SBP, the most common etiologies for SBP in cirrhosis are Enterobacter and Streptococcal species. Th e Aeromonas species are ubiquitous in fresh or sea water. Aeromonas caviae is never identified as etiology in cases of SBP. A patient, who had a history of liver cirrhosis related to chronic hepatitis B virus infection for 1 year, presented with diarrhea. He had diarrhea 1 week later returned from coastal city. He was hospitalized and treated with norfloxacin after 7 days of severe symptoms, including fever, abdominal distention, and diarrhea. Analysis of the ascitic specimen revealed a white-cell count of 4.42 × 109 cells/L with 88% neutrophils. Analysis of stool specimen showed a white-cell count of 60 cells per high-power field. Th e patient started the injection of cefriaxone at a dose of 4 g/d. However, the situation was not improved. Th ree days later, stool and ascitic fluid culture showed positive for Aeromonas caviae. Antibiotic susceptibility testing revealed that imipenem, meropenem, amikacin, and cefoperazone-sulbactam were highly sensitive to the Aeromonas caviae. However, the bacilli resisted to ceftriaxone, ceftazidime, ampicillin-sulbactam, levofloxacin, and sulfamethoxazole. Ceftriaxone was then switched to imipenem. The patient was fully recovered 14 days later. Aeromonas caviae is a rare pathogen of SBP in cirrhosis. It resists to third-generation of cephalosporin and fluroquinolone, which are of frequently used dependent on clinical experience. It needs a special attention.
Aeromonas caviae ; Anti-Infective Agents ; Ascitic Fluid ; Gram-Negative Bacterial Infections ; pathology ; Humans ; Leukocyte Count ; Liver Cirrhosis ; Male ; Microbial Sensitivity Tests ; Peritonitis ; microbiology ; pathology

A strain of Flavobacterium lindanitolerans isolated from a sick child's ascites was described. The 16S rRNA gene of the strain was 100% identical to that of Flavobacterium lindanitolerans which was first identified in India in 2008. It was first described that the isolate required X factor (Hemin) for growth in the optimal conditions of 37 °C with 5% CO(2). The isolate produced indole and H(2)S. It did not present hemolytic feature on blood agar.
Ascitic Fluid ; microbiology ; Child, Preschool ; Enterovirus A, Human ; isolation & purification ; Enterovirus Infections ; complications ; microbiology ; virology ; Fatal Outcome ; Flavobacteriaceae Infections ; complications ; microbiology ; virology ; Flavobacterium ; classification ; genetics ; isolation & purification ; Humans ; RNA, Ribosomal, 16S ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Selective intestinal decontamination (SID) with norfloxacin has been widely used for the prophylaxis of spontaneous bacterial peritonitis (SBP) because of a high recurrence rate and preventive effect of SID for SBP. However, it does select resistant gut flora and may lead to SBP caused by unusual pathogens such as quinolone-resistant gram-negative bacilli or gram-positive cocci. Enterococcus hirae is known to cause infections mainly in animals, but is rarely encountered in humans. We report the first case of SBP by E. hirae in a cirrhotic patient who have previously received an oral administration of norfloxacin against SBP caused by Klebsiella pneumoniae and presented in septic shock.
Administration, Oral ; Ampicillin/therapeutic use ; Anti-Bacterial Agents/therapeutic use ; Ascitic Fluid/microbiology ; Enterococcus/*isolation & purification ; Gram-Positive Bacterial Infections/complications/drug therapy/*microbiology ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Peritonitis/*diagnosis/drug therapy/microbiology ; Sepsis/*etiology

OBJECTIVETo investigate the levels of lipopolysaccharide binding protein (LBP) in serum and ascites of cirrhotic patients, and determine their diagnostic value for spontaneous bacterial peritonitis (SBP).

METHODSCirrhotic patients were divided into groups according to diagnosis of SBP, ascites without SBP, no ascites. To explore the significance of LBP in clinically suspect SBP cases, the ascites without SBP group was sub-divided into two groups according to the symptoms of abdominal pain or elevated white blood cell (WBC) count, and abdominal pain combined with elevated WBC count. Two control groups were composed of patients with intraperitoneal pus and a group of healthy, non-cirrhotic individuals. The LBP levels in serum and ascites were determined by enzyme-linked immunosorbent assay (ELISA). The ascites routine, ascites culture and albumin assay were carried out in the Second Affiliated Hospital of Chongqing Medical University. Data between the two groups were compared using the t-test or nonparametric test of independent samples, and the areas under the curve were compared using the Z test. Results The levels of LBP in serum and pus were significantly higher in the intraperitoneal pus group than in the cirrhosis group with ascites (P less than 0.01).

RESULTSThe level of serum LBP was significantly higher in the cirrhosis group with SBP than in the cirrhosis group without SBP but with ascites and the cirrhosis group with no ascites (P less than 0.01). There was no significant difference in the level of ascites LBP in the cirrhosis group with SBP and the cirrhosis group without SBP but with ascites (P more than 0.05). In the clinically suspect cases with SBP, the levels of LBP in serum and ascites were significantly higher than those in the cirrhosis group without SBP but with ascites (228.00 mug/ml vs. 80.95 mug/ml and 22.50 mug/ml vs. 11.45 mug/ml, P less than 0.05). Determination of serum LBP had a higher sensitivity than the determination of ascites LBP or ascites WBC.

CONCLUSIONGram-negative bacteria infection in the intra-abdominal cavity causes serum and body fluid levels of LBP to increase significantly. Patients with cirrhosis complicated with SBP have significantly elevated levels of serum LBP. The serum and ascites LBP levels are significantly elevated in SBP patients with suspected clinical diagnosis. Measurements of both the serum LBP and ascites LBP may have diagnostic value for SBP.

Acute-Phase Proteins ; metabolism ; Adult ; Aged ; Ascites ; diagnosis ; microbiology ; Ascitic Fluid ; chemistry ; Bacterial Infections ; complications ; diagnosis ; Carrier Proteins ; blood ; metabolism ; Case-Control Studies ; Female ; Humans ; Liver Cirrhosis ; complications ; microbiology ; Male ; Membrane Glycoproteins ; blood ; metabolism ; Middle Aged ; Peritonitis ; complications ; diagnosis ; microbiology

OBJECTIVETo explore the relationship of ascitic bacterial DNA and plasma endotoxin, intestinal permeability, gut flora in cirrhotic patients.

METHODSFifty-five decompensated cirrhotic patients with ascites were included in the study. A paracentesis was performed for every patient at admission and ascites fluid was collected for bacterial DNA isolation and amplification, plus other routine studies and cultured for aerobic and anaerobic bacteria. Plasma endotoxin, intestinal permeability, and gut flora were studied on the day following admission. Blood from the patients was obtained for routine hematologic, biochemical, and coagulation studies. Thirty healthy subjects were studied as normal controls.

RESULTSNo bacteria were found in the ascites cultures in the 55 patients, but bacterial DNA was found in 19 (34.55%). Compared with the bacterial DNA negative group, the bacterial DNA positive group showed a significantly lower level of PTA (t= -3.184, P=0.002), a higher Child-Pugh score (t=3.224, P=0.002) and a higher quantity of WBC in their ascitic fluid (t=4.088, P=0.001). Compared with normal controls, cirrhotic patients showed significantly higher levels of plasma endotoxin (t=13.705, P=0.000), lactulose/mannitol (L/M, t=28.568, P=0.000) in urine, and the quantity of enteric bacilli (t=2.912, P=0.005); the quantity of their intestinal bifidobacteria was significantly lower (t= -3.669, P=0.000). The variables correlative with the presence of bacterial DNA were the quantities of enteric bacilli (P=0.007) and PTA (P=0.011).

CONCLUSIONIntestinal bacterial overgrowth plays a key role in the pathogenesis of ascitic bacterial translocation (ABT) in cirrhotic patients. ABT is correlated with the degrees of the liver disease.

Adult ; Aged ; Aged, 80 and over ; Ascites ; etiology ; microbiology ; Ascitic Fluid ; microbiology ; Bacterial Translocation ; DNA, Bacterial ; isolation & purification ; Enterobacteriaceae ; genetics ; isolation & purification ; Female ; Humans ; Liver Cirrhosis ; complications ; microbiology ; Male ; Middle Aged

BACKGROUND/AIMS: Spontaneous bacterial peritonitis (SBP) is one of the potentially life-threatening complications for patients with liver cirrhosis, and it has a mortality rate of over 20%. Early diagnosis of SBP and immediate use of an adequate antibiotic therapy are very important for achieving a better prognosis. The aim of our study was to assess the usefulness of reagent strips for making the rapid diagnosis of SBP. METHODS: A diagnostic paracentesis procedure was performed upon hospital admission in 257 cirrhotic patients (187 males, 70 females; mean age: 54 years) with ascites. Each fresh sample of ascitic fluid was tested using a reagent strip, and the result was scored as 0, 1+, 2+ or 3+. The leukocyte count, polymorphonuclear cell count, blood bottle culture, and chemistry of ascites were also done. RESULTS: We diagnosed 79 cases of SBP and 2 cases of secondary bacterial peritonitis by means of the polymorphonuclear cell count and the classical criteria. When a reagent strip result of 3+ was considered positive, the test's sensitivity was 86% (70 of 81), the specificity was 100% (176 of 176), and the positive predictive value was 94%. Furthermore, when a reagent strip result of 2+ or more was considered positive, the test sensitivity was 100% (81 of 81), the specificity was 99% (174 of 176), and negative predictive value was 99%. CONCLUSIONS: The use of reagent strips is a very sensitive and specific tool for the rapid diagnosis of SBP in cirrhotic patients. A positive result should be an indication for empirical antibiotic therapy, and a negative result may be useful as a screening test to exclude SBP.
Aged ; Ascitic Fluid/chemistry/cytology ; Bacterial Infections/*diagnosis/microbiology ; English Abstract ; Female ; Humans ; Liver Cirrhosis/complications ; Male ; Middle Aged ; Peritonitis/*diagnosis/etiology/microbiology ; Predictive Value of Tests ; *Reagent Strips ; Sensitivity and Specificity

PURPOSE: This study focused on the effect of immuno-compromising conditions on the clinical presentation of severe listerial infection. PATIENTS AND METHODS: Nine human listeriosis cases seen from 1991-2002 were reviewed. All adult patients, from whose blood, peritoneal fluid or cerebrospinal fluid (CSF) the L. monocytogenes was isolated, were included in this retrospective study. RESULTS: Listeriosis presented as primary sepsis with positive blood cultures in 5 cases and meningitis with positive CSF cultures in 4 cases. All of these patients had at least one underlying disease, most commonly, hematologic malignancy, diabetes mellitus, amyloidosis and hepatic cirrhosis; 55.6% had received immunosuppressive or corticosteroid therapy within a week before the onset of listeriosis. The patients were adults with a mean age of 60 years. Fever, night sweats, chills and lethargy were the most common symptoms; high temperature (> 38 degrees C), tachycardia, meningeal signs and poor conditions in general were the most common findings on admission. The mortality rate was 33.3% and was strictly associated with the severity of the underlying disease. Mortality differences were significant between sepsis (20%) and meningitis (50%) patients. CONCLUSION: Listeriosis as an uncommon infection in our region and that immuno- suppressive therapy is an important pre-disposing factor of listeriosis. Sepsis and meningitis were more common in this group of patients and had the highest case-fatality rate for food-borne illnesses.
Adrenal Cortex Hormones/therapeutic use ; Adult ; Aged ; Ascitic Fluid/microbiology ; Female ; Humans ; Immunosuppressive Agents/*therapeutic use ; Listeria Infections/blood/cerebrospinal fluid/*drug therapy ; Listeria monocytogenes/*drug effects/growth & development/isolation & purification ; Male ; Meningitis, Bacterial/blood/cerebrospinal fluid/*drug therapy ; Middle Aged ; Retrospective Studies ; Sepsis/blood/cerebrospinal fluid/*drug therapy ; Treatment Outcome

OBJECTIVETo explore the clinical value of serum procalcitonin (PCT) for predicting spontaneous bacterial peritonitis (SBP) in end-stage liver diseases.

METHODSThe clinical data of 362 ascitic inpatients with end-stage liver diseases who had underwent serum PCT assay in our department from March 2011 to June 2013 were analyzed retrospectively. These patients were then divided into SBP group (n=178) and non-SBP group (n=184). The dynamic changes of the PCT values upon admission and after antibiotic treatment were compared. The receiver operating characteristic curve was drawn to identify the optimal cut-off value of serum PCT in diagnosing SBP.

RESULTSThe positive rate of bacteria culture in ascites was only 4.6% (4/87) in SBP group. The median value of serum PCT was 0.73 and 0.15 ng/ml in SBP group and non-SBP group (Z=-11.9, U=0.000), respectively, before antibiotic treatment. In the SBP group, the median value of serum PCT was 1.73 ng/ml in 13 patients with positive culture findings, which was higher than the overall median value in SBP group. Among patients who were responsive to the antibiotic therapy, the median values of serum PCT were 0.40(n=46), 0.32(n=19), and 0.33 ng/ml(n=25), respectively, 3, 5, and 7 days after the effective antibiotics treatment, which were significantly lower than the pre-treatment levels [0.86(Z=-5.91, U=0.000), 0.72(Z=-3.10, U=0.002), and 0.79 ng/ml(Z=-4.37, U=0.000), respectively]. ROC analysis showed that a serum PCT value of more than 0.462 ng/ml had a sensitivity of 83.7% and a specificity of 94.9%(AUC:0.95, 95%CI:0.93-0.97, P=0.00) in diagnosing SBP in patients with end-stage liver diseases.

CONCLUSIONSAscitic fluid positive rate is low in SBP patients. Serum PCT is a sensitive and specific marker for predicting peritoneal bacteria infection in end-stage liver disease patients with ascites. Higher serum PCT can be expected in these patients with heavier infections, it can also be used to evaluate the effectiveness of anti-bacteria therapies.

Adult ; Aged ; Ascitic Fluid ; microbiology ; Bacterial Infections ; complications ; diagnosis ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Female ; Humans ; Liver Diseases ; complications ; Male ; Middle Aged ; Peritonitis ; complications ; diagnosis ; Protein Precursors ; blood ; Retrospective Studies ; Sensitivity and Specificity