ABSTRACT
<p><b>OBJECTIVE</b>To establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.</p><p><b>METHODS</b>A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on.</p><p><b>RESULTS</b>The linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.</p><p><b>CONCLUSION</b>Established ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.</p>
Subject(s)
Humans , Antibodies, Monoclonal , Enzyme-Linked Immunosorbent Assay , Methods , Membrane Proteins , Blood , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection.</p><p><b>METHODS</b>E. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast.</p><p><b>RESULTS</b>A total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79.</p><p><b>CONCLUSION</b>CTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.</p>
Subject(s)
Humans , Bacteremia , Microbiology , Cross Infection , Microbiology , Drug Resistance, Bacterial , Escherichia coli , Genetics , Escherichia coli Infections , Microbiology , Escherichia coli Proteins , Genetics , Genotype , Hospitalization , Liver Cirrhosis , Therapeutics , Microbial Sensitivity Tests , beta-Lactamases , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To characterize the clinical, laboratory, imaging and pathological features of primary sclerosing cholangitis (PSC) and investigate the impact of ursodeoxycholic acid (UDCA) therapy on patient prognosis.</p><p><b>METHODS</b>The medical records of 22 patients diagnosed with PSC between 2002 and 2011 were retrospectively reviewed. The PSC diagnosis had been made in patients with suspect biochemical abnormalities following evaluation by magnetic resonance cholangiopancreatography (MRCP) and/or endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic cholangiography (PTC). Fibrosis and inflammation were assessed by immunohistochemical analyses of tissue biopsies. Outcome of patients treated with UDCA (13-15 mg/kg/day, oral) were compared to that of patients without UDCA treatment by the X2 or corrected X2 tests.</p><p><b>RESULTS</b>Among the 22 PSC patients, the majority was male (n=15) and presented with fatigue, dark urine, and body weight loss (n=15). Four cases had ulcerative colitis. At admission, all 22 cases showed elevated levels of alkaline phosphatase[ALP: (348+/-184) U/L], 19 cases showed elevated alanine aminotransferase [ALT: (94.0+/-67.0) U/L] and aspartate aminotransferase [AST: (98.0+/-67.0) U/L], and 15 cases showed elevated levels of total bilirubin (99.0+/-115.0) mumol/L and direct bilirubin (74.4+/-92.4 mumol/L. ERCP examination showed segmental intrahepatic bile duct stenosis with expansion, and stiff and enlarged gallbladder bile ducts, but unclear findings for the common bile ducts and pancreatic ducts. MRCP showed beading of the intrahepatic bile duct, stiffness of the bile duct wall, and dilation of the common bile duct. Fibrosis and inflammation were observed in the bile ducts, along with hyperplasia and the typical features of "onion skin" fibrosis and fibrous obliterative cholangitis. Five of the 10 patients treated with UDCA improved, and seven of the 12 patients in the non-UDCA treatment group improved. There was no statistically significant difference in outcome between the groups (paired X2=0.333, corrected X2=0.083, P more than 0.05).</p><p><b>CONCLUSION</b>PSC patients were predominantly male and the common clinical manifestations were fatigue, dark urine, and body weight loss. At admission, serum biochemical indicators of cholangitis were increased significantly and subsequent imaging studies confirmed the suspected diagnosis by showing obvious characteristic changes. UDCA treatment did not significantly improve patient prognosis.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Cholangiography , Methods , Cholangiopancreatography, Endoscopic Retrograde , Cholangitis, Sclerosing , Diagnostic Imaging , Pathology , Retrospective StudiesABSTRACT
<p><b>OBJECTIVE</b>Assessment of detection of IgM antibodies for human enterovirus 71 (EV 71) in early diagnosis for the hand, foot and mouth disease (HFMD).</p><p><b>METHOD</b>The sera and throat swabs from 38 patients which were clinical diagnosis as HFMD, were continuous daily collected in our hospital in 2010. These specimens were detected by EV 71 IgM antibodies assay, real time RT-PCR methods for EV 71 and Enterovirus.</p><p><b>RESULTS</b>Among 38 HFMD patients, the cumulative positive rates of EV 71 IgM antibodies were: 60.5% on day 1, 71.1% on day 2, 81.5% in the first 3-4 days, 92.1% on day 5, 92.1% on day 6, and the positive rate of nucleic acid detected by the real time RT-PCR for EV 71 and Enterovirus were 60.5%, 73.6%.</p><p><b>CONCLUSION</b>The positive rate of EV 71 IgM antibodies in the hand, foot and mouth disease just can occur on day 1, and reach to peak on day 5, which can be used as one of indicators of early diagnosis of hand, foot and mouth disease.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Antibodies, Viral , Blood , Allergy and Immunology , Early Diagnosis , Enterovirus A, Human , Allergy and Immunology , Hand, Foot and Mouth Disease , Diagnosis , Allergy and Immunology , Virology , Immunoglobulin M , Blood , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To verify a new kit of "universal and novel influenza A (H1N1) virus nucleic acid double-detection methods (PCR-fluorescence probe)".</p><p><b>METHODS</b>150 cases of throat swab specimens were collected consecutively. After RNA was extracted, the specimens were detected by the verified kit. At the same time, the same specimens were detected by Real-time PCR diagnostic kit from Beijing CDC as the control. The data were analysed by the Kappa in agreement and by McNemar chi2 in difference test.</p><p><b>RESULTS</b>The consistency rate of the verified kit and the Beijing CDC kit was universal primer M 97.33%, H1N1 98.67% respectively. The Kappa test and McNemar chi2 test showed that two methods had a higher consistency. Compared to the CDC kit, the "false negative rate" and "false-positive rate"of double-check kit were lower.</p><p><b>CONCLUSION</b>The kit of "universal and novel influenza A (H1N1) virus nucleic acid double-detection methods (PCR-fluorescence probe)" from Shanghai Kehua Bio-Engineering Co., Ltd can be used to detect influenza A and novel influenza A (H1N1).</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , DNA, Viral , Influenza A Virus, H1N1 Subtype , Genetics , Polymerase Chain Reaction , Methods , Reagent Kits, DiagnosticABSTRACT
<p><b>OBJECTIVE</b>To analysis the clinical and laboratory characteristics of Patients infected with new influenza A (HIN1) virus.</p><p><b>METHODS</b>All cases with new influenza A (H1N1) confirmed on polymerase chain reaction assay on throat swabs. There were included in a prospective evaluation of clinical characteristics, laboratory results, treatment and overcome of new influenza A (H1N1).</p><p><b>RESULTS</b>There were 35 patients in the epidemic. Clinical illness developed within a mean of 1.7 days. Fever occurred in 97.1%, sore throat 65.7% cough 51.4%, headache 28.6%, and myalgia 31.4%. All patients were treated with oseltamivir lasted 5 days. The mean duration of viral shedding was 4.5 days. All were cured and left hospital after day 7.</p><p><b>CONCLUSION</b>It was infected by new influenza A (H1N1) typically in this epidemic.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Disease Outbreaks , Influenza A Virus, H1N1 Subtype , Virulence , Influenza, Human , Diagnosis , Drug Therapy , Virology , Oseltamivir , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To study the status of beta-lactamase produced by multiresistant Aeromonas selected from cirrhosis patients to provide reference for treatment and reduce resistance and control spreading.</p><p><b>METHODS</b>Four multiresistant Aeromonas strains isolated from serious liver cirrhosis patients from the No. 302 hospital. The TEM resistant genes were detected by PCR and agarose gel electrophoresis.</p><p><b>RESULTS</b>Three TEM-1 positive strains were detected from four multiresistant Aeromonas isolates consisting of one Aeromonas sobria and three Aeromonas hydrophila isolated from blood and ascites. This was further confirmed by gene sequencing. The multiresistance to antibiotics was higher in four Aeromonas isolates. All strains tested were resistant to ampicillin, cefazolin and cefmetazole.The cirrhosis patients who suffered from Aeromonas infection had poor prognosis and had mortality rate of 3/4.</p><p><b>CONCLUSION</b>The beta-lactamase TEM-1 resistant genes was detected in clinical multiresistant Aeromonas strain isolated from serious cirrhosis patients.The results suggested that TEM-1 was the main resistance mechanism of Aeromonas strain and was reduced by adding enzyme inhibitor.</p>