[Quantitative analyses of intrahepatic HBV cccDNA and serum HBsAg in 54 patients with chronic hepatitis B].

OBJECTIVE: To quantitatively detect intrahepatic HBV covalently closed circular DNA (cccDNA) and serum HBsAg; and to analyze the relationship between the two parameters and with serum HBV DNA level. METHODS: Intrahepatic cccDNA (copies/cell) was quantitated by plasmid-safe ATP-dependent Danes (PSAD) digestion in combination with rolling circle amplification and gap-spanning selective real-time PCR assay using formalin fixed paraffin-embedded liver biopsy samples. HBsAg was measured by chemiluminescence's reagent manufactured by Abbott Company using sera sampled at time-point of liver biopsy. RESULTS: Intrahepatic cccDNA level was positively correlated with serum HBsAg level (r = 0.459, P < 0.001), but not correlated with serum HBV DNA level. Serum HBsAg level was positively correlated with serum HBV DNA level (r = 0.328, P = 0.015), and reversely correlated with HBV replicative efficiency defined as the ratio of serum HBV DNA to cccDNA (r = -0.373, P = 0.005). CONCLUSION: In patients with chronic hepatitis B, intrahepatic cccDNA level is correlated with serum HBsAg level. The two parameters combined with serum HBV DNA may comprehensively reflect HBV replication activity and help evaluation of antiviral therapeutic efficacy.

Multicentre evaluation of the Elecsys hepatitis B surface antigen II assay for detection of HBsAg in comparison with other commercially available assays.

Screening for hepatitis B surface antigen (HBsAg) demands highly sensitive and specific immunoassays with the ability to detect clinically relevant surface gene mutants-the presence of which is an increasing concern and can compromise test results. The objective of the study was to compare the clinical and technical performance of the fully automated Elecsys HBsAg II assay with other widely used HBsAg immunoassays in China. This was a multicentre study in which eight Chinese laboratories compared the Elecsys HBsAg II assay with the Architect HBsAg assay, AxSYM HBsAg assay, or generic microtitre plate (MTP) enzyme-linked immunosorbent assays (manufactured in China) against preselected samples, including recombinant surface gene mutants, and routine clinical samples. Elecsys HBsAg II was the most sensitive assay for detecting positive results in seroconversion samples: up to 14, 11, and 22 days earlier than the Architect, AxSYM, and MTP HBsAg assays, respectively. It also detected all 211 preselected HBsAg-positive specimens and all 13 recombinant HBsAg mutants; the AxSYM and MTP HBsAg assays failed to detect 3 and 10 mutant samples, respectively. Sensitivity was 100% for Elecsys HBsAg II with routine samples, compared with 99.1, 98.9, and 95.2% for the Architect, AxSYM, and MTP HBsAg assays, respectively. In conclusion, it was demonstrated that the Elecsys HBsAg II assay is suitable for routine HBsAg screening in China.

[The distribution and antimicrobial resistance tendency of pathogens associated with diarrhea in Beijing].

OBJECTIVE: To monitor the constituents and resistant tendency of bacterial pathogens isolated from diarrheal patients in our hospital form 1994 to 2005 to offer the basis for guiding epidemiologic study, vaccination research and clinical treatment. METHODS: Enteric pathogenic bacteria were cultured and identified to species, group and serotype with biochemical and serologic methods and the susceptibility of bacteria to antimicrobial agents were tested. RESULTS: Enteric pathogenic bacteria were isolated predominantly in male patients and mainly in children and youngsters. It reached a peak from July to September every year. Shigella spp. (75.11%) was the most frequently isolated pathogens and followed by Vibrio spp. (12.7%), Salmonella spp. (6.28%), Aeromonas spp. (4.43%) and Escherichia coli (1.25%). During the period from 1994 to 2005, diarrheal pathogens had a trend of decrease especially Shigella spp. and Salmonella spp. Of the 6329 isolates of Shigella spp., 75.62% was S. flexneri and S. sonnei, S. dysenteriae and S. boydii constituted 23.98%, 0.22% and 0.01% respectively. The sensitivity of different species, group or serotype to different antimicrobial agents was not the same. S. flexneri and Aeromonas spp. were highly resistant to most of antibiotics. However, S. sonnei and Vibrio spp. had good susceptibility to antibiotics tested except trimethoprim/sulfamethoxazole and ampicillin. CONCLUSION: There are many species and serotypes of enteric pathogenic bacteria causing infective diarrhea and the distribution changes gradually in Beijing. The resistance rate of enteric pathogenic bacteria to antibiotics is not the same in different species and serotypes, so strict surveillance is always needed.

Performance verification and comparison of TianLong automatic hypersensitive hepatitis B virus DNA quantification system with Roche CAP/CTM system.

AIM: To investigate and compare the analytical and clinical performance of TianLong automatic hypersensitive hepatitis B virus (HBV) DNA quantification system and Roche CAP/CTM system. METHODS: Two hundred blood samples for HBV DNA testing, HBV-DNA negative samples and high-titer HBV-DNA mixture samples were collected and prepared. National standard materials for serum HBV and a worldwide HBV DNA panel were employed for performance verification. The analytical performance, such as limit of detection, limit of quantification, accuracy, precision, reproducibility, linearity, genotype coverage and cross-contamination, was determined using the TianLong automatic hypersensitive HBV DNA quantification system (TL system). Correlation and Bland-Altman plot analyses were carried out to compare the clinical performance of the TL system assay and the CAP/CTM system. RESULTS: The detection limit of the TL system was 10 IU/mL, and its limit of quantification was 30 IU/mL. The differences between the expected and tested concentrations of the national standards were less than ± 0.4 Log10 IU/mL, which showed high accuracy of the system. Results of the precision, reproducibility and linearity tests showed that the multiple test coefficient of variation (CV) of the same sample was less than 5% for 102-106 IU/mL; and for 30-108 IU/mL, the linear correlation coefficient r2 = 0.99. The TL system detected HBV DNA (A-H) genotypes and there was no cross-contamination during the "checkerboard" test. When compared with the CAP/CTM assay, the two assays showed 100% consistency in both negative and positive sample results (15 negative samples and 185 positive samples). No statistical differences between the two assays in the HBV DNA quantification values were observed (P > 0.05). Correlation analysis indicated a significant correlation between the two assays, r2 = 0.9774. The Bland-Altman plot analysis showed that 98.9% of the positive data were within the 95% acceptable range, and the maximum difference was -0.49. CONCLUSION: The TL system has good analytical performance, and exhibits good agreement with the CAP/CTM system in clinical performance.

[Identification of circulating plasmacytoid dendritic cells in chronic HBV-infected patients].

OBJECTIVE: To investigate the features of circulating plasmacytoid dendritic cells (pDCs) in patients with chronic hepatitis B for a better understanding of the immunopathogenesis of HBV infection. METHODS: Fresh blood samples were collected from 20 patients with chronic hepatitis B (CHB) and from 15 healthy individuals who served as controls. pDCs were isolated from peripheral blood mononuclear cells (PBMCs) using immunomagnetic assay and detected by flow cytometry. Fresh PBMCs and isolated pDCs were stimulated in vitro using CpG ODN2216. The supernatants were measured for IFNa production using ELISA. RESULTS: The peripheral pDCs frequency in CHB patients (0.192%+/-0.110%) was markedly lower than that in the healthy controls (0.287%+/-0.142%). After being pulsed with CpG ODN2216, the isolated pDCs produced lower levels of IFNa and expressed lower levels of CD80 and CD40 in the CHB patients when compared to those of the healthy controls. The level of IFNa was (972.6+/-705.5) pg/ml in the patients and (3 142.9+/-1 292.2) pg/ml in the controls. Moreover, the pDCs frequency was reversely correlated with serum ALT levels in these HBV infected patients. CONCLUSION: The reduced number and impaired function of circulating pDCs in patients with CHB may be related to their disease progression.

Autoantibodies in Chinese patients with chronic hepatitis B: prevalence and clinical associations.

AIM: To investigate the prevalence of autoantibodies and their associations with clinical features in Chinese patients with chronic hepatitis B (CHB). METHODS: A total of 325 Chinese patients with CHB were enrolled in this retrospective, hospital-based study. Patients with chronic hepatitis C (CHC), autoimmune hepatitis (AIH), or primary biliary cirrhosis (PBC) were included, with healthy donors acting as controls. A panel of autoantibodies that serologically define AIH and PBC was tested by indirect immunofluorescence assay and line immunoassay. The AIH-related autoantibody profile included homogeneous anti-nuclear antibodies (ANA-H), smooth-muscle antibodies, anti-liver kidney microsome type 1, anti-liver cytosolic antigen type 1, and anti-soluble liver antigen/liver pancreas; the PBC-related antibodies were characterized by ANA-nuclear dots/membranous rim-like, anti-mitochondrial antibodies-M2 (AMA-M2), anti-BPO (recombinant antigen targeted by AMA-M2), anti-Sp100, anti-promyelocytic leukemia protein (anti-PML), and anti-gp210. The dichotomization of clustering was used to unequivocally designate the AIH or PBC profiles for each case. Anti-Ro52 antibodies were also tested. RESULTS: The prevalence of any autoantibody in CHB amounted to 58.2%, which was similar to the 66.2% prevalence in CHC, significantly higher than the 6.7% in the healthy controls (P < 0.001), and lower than the 100% found in AIH and PBC (P = 0.004 and P < 0.001, respectively). There were more anti-PML and anti-gp210 antibodies among the CHB patients than the CHC patients (11.1% vs 0%, P = 0.003; 12.6% vs 0%, P < 0.001, respectively). The prevalence and titer of AMA, anti-BPO, anti-PML, and anti-gp210 were higher in PBC than in those with CHB. Among the CHB patients, the prevalence of ANA, especially ANA-H, was significantly lower in patients with compensated and decompensated cirrhosis compared with patients without cirrhosis. Thirty-eight cases of hepatocellular carcinoma (HCC) in CHB showed a significant difference compared with non-HCC patients in the prevalence of anti-PML (0% vs 12.5%, P = 0.013). Dichotomization of the autoantibodies revealed that the PBC profile was more prevalent in patients with CHB than in those with CHC, and that it was strongly correlated with both compensated and decompensated cirrhosis. In contrast, the prevalence of the AIH profile was significantly higher in non-cirrhosis patients with CHB than in those with compensated cirrhosis (18.5% vs 8.2%, P = 0.039). Moreover, the AIH profile was also closely associated with hepatitis B e-antigen positivity. CONCLUSION: ANA-H could be an indicator of early-stage CHB. Dichotomizing the autoantibody profiles revealed that the PBC profile is strongly associated with cirrhosis in CHB.

[The clinical features and prognosis of Aeromonas septicaemia in hepatic cirrhosis: a report of 50 cases].

OBJECTIVE: Aeromonas septicaemia complicating cirrhosis is not a common infectious disease. To enhance the knowledge in this aspect, we analysed the clinical features, pathogenetic factors, resistance, treatment and prognosis of Aeromonas septicaemia in 50 cases of hepatic cirrhosis. METHODS: We cultured the bacteria from infected patients with BacT/Alert120 automation instrument made in AKsu and identified the bacteria with the Vitek-AMS60 made in Biomerieux company. We then tested the susceptibility of Aeromonas to 13 antimicrobial agents. RESULTS: A total of 50 cases of Aeromonas septicaemia occurred in severe hepatic cirrhosis. The majority of them had severe complications. Aeromonas hydrophila was the most common species isolated (52.0%). Nosocomial infection was the predominant way of infection. The major clinical manifestations of Aeromonas septicaemia were fever (100%), chill (64.0%), abdominal pain (60.0%), diarrhoea (32.0%) and shock (24.0%). The susceptive rate of Aeromonas to third generation cephalosporin, quinolones and aminoglycoside antibiotics was more than 80%. The cure rate and mortality attributed to Aeromonas septicaemia after treating with third generation cephalosporin, levofloxacin and their combination were 64.3%, 75.0%, 57.1% and 28.6%, 12.5%, 35.7% respectively. CONCLUSION: Aeromonas septicaemia tends to befall patients with severe hepatic cirrhosis and causes a rapidly fatal outcome. Aeromonas should be considered an important pathogen for septicemia in patients with liver cirrhosis. It is suggested that we should emphasize the clinical features and laboratory diagnosis so as to have proper antimicrobial treatment.

[Analysis of severe acute respiratory syndrome in Beijing].

OBJECTIVE: To study the clinical, laboratory, and radiologic features of 34 cases of severe acute respiratory syndrome (SARS) in Beijing. METHODS: All patients were admitted to the isolation wards. Their demographic, clinical, laboratory, and radiologic characteristics were analyzed. Univariate and multivariate analyses were performed. RESULTS: Eight patients came from a family, and 15 patients were medical staff. The mean age of patients was (33.4 +/- 13.4) years. The latent period varied from 2 to 14 days (median 4 days). The most common symptoms were fever (100%), palpitation (91.7%), myalgia (79.2%), headache (70.8%), diarrhea (73.9%) and cough (58.3%). The mean leucocyte count was (4.6 +/- 1.4) x 10(9)/L, and the mean lymphocyte ratio was 0.27 +/- 0.11. 68.4% of the patients had lymphopenia (absolute lymphocyte count < 1.3 x 10(9)/L). Other common findings included elevated levels of serum alanine aminotransferase, lactate dehydrogenase and erythrocyte sedimentation (76.2%, 28.6% and 47.8%, respectively), and decreased levels of serum iron and albumin (63.2% and 47.8%, respectively). Thirty-two cases had abnormal chest radiographs. In 2 cases in whom typical lung opacities could not be found on the initial plain chest radiographs, thoracic CT proved to be useful. Postmortem examination of 1 patient revealed marked edema with foci of hemorrhage and hyaline membrane formation in the lungs, hemorrhage necrosis and a obvious decline of cells in lymph glands. In a multivariate analysis (Stata 7.0), the independent predictor of an adverse outcome was advanced age (odds ratio per decade of life, 1.6; 95% CI, 1.08 to 2.63; P = 0.007). CONCLUSIONS: Fever, lymphopenia, low serum iron and chest radiograph are helpful to diagnose SARS early; age is the independent predictor of an outcome.

[The investigantion of comparative experiments for different clinical laboratory instruments].

OBJECTIVE: To estimate the consistency of two VITROS 3600 chemiluminescent analyzers according to the requirement of ISO15189. METHODS: Verification tests were made for precision and accuracy of anti-HCV in two instruments. While 40 serum samples including Anti-HCV negative (10 cases) , positive (10 cases) , and weakly positive (20 cases). and the test results were statistical analised. RESULTS: Two instruments negative and positive control samples intra-batch precision and coefficients of variation were 5% , 4% and 7. 14% , 7. 23% , inter-batch precision and coefficients of variation were 9. 47% , 7. 7% and 8.04%, 7. 6%, are less than requirement CV (15%) by ISO15189. The accuracy of two instrument were 100% , The test results of the control samples showed no significant difference (P < 0. 05). The correlation analysis of the test results of clinical samples R2 =0. 9984, with good consistency. CONCLUSION: Test results of two Vitros 3600 has good consistency and comparability.

[Distribution of genotypes in ESBLs producing E. coli strains isolated from posthepatitic cirrhosis' patients with bloodstream infection].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Dynamic analysis of the lymphocyte subsets in HCV children with different genotypes during treatment].

OBJECTIVE: To discuss the changes of lymphocyte subsets in HCV children with different genotypes during treatment with pegylated interferon alfa-2b and ribavirin. METHODS: The genotype of 45 HCV infected children were identified by real time PCR. The lymphocyte subsets were dynamically detected by BD FACSCalibur flow cytometer with four color MultiTEST IMK Kit during the treatment. RESULTS: For the children with 1b genotype, after 24 weeks, the CD4+ T cells were higher than pre-treatment (P < 0.05). For the children with 2a genotype, after 12 weeks and after 24 weeks, the CD3+ T cells and CD4+ T cells significantly increased while the NK cells decreased than pre-treatment (P < 0.05). CONCLUSIONS: The lymphocyte subsets of HCV children with 2a genotype were different from 1b genotype during trentment with pegylated interferon alfa-2b and ribavirin.

Granulocyte-colony stimulating factor therapy improves survival in patients with hepatitis B virus-associated acute-on-chronic liver failure.

AIM: To evaluate the safety and efficacy of granulocyte-colony stimulating factor (G-CSF) therapy in patients with hepatitis B virus (HBV)-associated acute-on-chronic liver failure (ACLF). METHODS: Fifty-five patients with HBV-associated ACLF were randomized into two groups: the treatment group and the control group. Twenty-seven patients in the treatment group received G-CSF (5 µg/kg per day, six doses) treatment plus standard therapy, and 28 patients in the control group received standard therapy only. The peripheral CD34(+) cell count was measured consecutively by flow cytometry. Circulating white blood cell count, biochemical parameters, and other clinical data of these patients were recorded and analyzed. All patients were followed up for a period of 3 mo to evaluate the changes in liver function and survival rate. RESULTS: The peripheral neutrophil and CD34(+) cell counts in the G-CSF group increased on day 3 from the onset of therapy, continued to rise on day 7, and remained elevated on day 15 compared to those of the control group. Child-Turcotte-Pugh score of patients in the treatment group was improved on day 30 from the onset of G-CSF therapy, compared to that in the controls (P = 0.041). Model for End-Stage of Liver Disease score of patients in the treatment group was improved on day 7 (P = 0.004) and remained high on day 30 from the onset of G-CSF therapy (P < 0.001) compared to that in controls. After 3 mo of follow-up observation, the survival rate in the treatment group (48.1%) was significantly higher than that in the control group (21.4%) (P = 0.0181). CONCLUSION: G-CSF therapy promoted CD34(+) cell mobilization in patients with HBV-associated ACLF, and improved the liver function and the survival rate of these patients.

[Establishment of a purificatory method for alpha-fetoprotein variant by affinity adsorption].

OBJECTIVE: To establish a purificatory method of alpha-fetoprotein variant (AFP-L3) based on microspincolumn with lens culinaris agglutinin (LCA). METHODS: LCA was isolated by ammonium sulfate precipitation method from lens culinaris. AFP-L3 affinity adsorption microspincolumns which were made from LCA coupled with activated Sepharose 4B were prepared. By adding into the centrifuge column, serum was absorbed and eluted to purify AFP-L3. The results of purified AFP-L3 detection of 10 cases AFP positive sera by electro-chemiluminescence immunoassay were compared with traditional crossed affinity immunoelectrophoresis. RESULTS: 8 of 10 cases AFP-L3 concentration were greater than 5 ng/ml in purified sera. Six cases show positive reaction in affinity immune cross electrophoresis experiment. CONCLUSION: Successfully established purification method of AFP-L3 by affinity absorption based on microspincolumn. The method was more conducive to clinical laboratory applications due to its high sensitive and easy operation.

[Cloning and expressing of golgi protein73 gene fragment and preparation of monoclonal antibodies against the recombinant protein].

OBJECTIVE: To clone and express human Golgi glycoprotein73 protein, and prepare the monoclonal antibody (mAb) against the protein. METHODS: GP73 gene was amplified from HepG2 cells by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-GP73 and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot was used to detect specificity of mAbs. RESULTS: The prokaryotic plasmid expressing the recombinant protein was constructed, and the GP73 recombinant protein was expressed and purified. Five hybridoma cell lines that secreted anti-GP73 mAbs were obtained. 2 of 5 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with GP73 protein. CONCLUSION: The GP73 recombinant protein is highly purified and has strong antigenicity. The anti-GP73 mAbs were prepared successfully.

[Cloning and expressing of tissue inhibitor of metalloproteinases I gene fragment and preparation of monoclonal antibodies against the recombinant protein].

OBJECTIVE: To prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein. METHODS: TIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot were used to detect specificity of mAbs. RESULTS: The prokaryotic plasmid expressing the recombinant protein was constructed, and the TIMP-I recombinant protein was expressed and purified. Four hybridoma cell lines that secreted anti-TIMP-I mAbs were obtained. 3 of 4 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with TIMP-I protein. CONCLUSION: The TIMP-I recombinant protein is highly purified and has strong antigenicity. The anti- TIMP-I mAbs were prepared successfully.

[Analysis of hemagglutination inhibition antibody level in patients with influenza A H1N1].

OBJECTIVE: To understand the hemagglutination inhibition antibody level in patients with influenza A H1N1. METHODS: Sera from 28 patients with influenza A H1N1 at different time points after illness onset were collected and measured by hemagglutination inhibition assay. RESULTS: The serum hemagglutination inhibition antibody titers at 1, 5, 15, 22, 37, 49 and 58 days after illness onset were 5.36, 9.39, 39.02, 57.99, 137.92, 55.19 and 57.99 respectively. The top geometric mean titer of hemagglutination inhibition antibody was 148.55. The antibody seroconversion rate and seroprotection rate were occurred in 96.4% (27/28) of patients. CONCLUSION: The patients with influenza A H1N1 have effective immune response.

[Establishment of confirmatory test for suspicious hepatitis B surface antigen positive samples].

OBJECTIVE: Establish a confirmatory test based on ELISA, and use to verify the authenticity of HBsAg weak positive samples, pick and get rid of the false result, and avoid the mistake diagnosis. METHOD: The particles (reagent A) coated by streptavidin and biotinylated HBsAb (reagent B) were mixed in different proportions, then neutralized with serum whose the COI of HBsAg > 20 by ELISA in order to identify the activity of HBsAb in confirmatory reagent. 30 pieces of HBsAg weak positive serum neutralized with the confirmatory reagent, the serum were considered to be positive if rate of decline of HBsAg COI > 50%. The results were compared to Roche confirmatory Kit. RESULT: Confirmatory reagent was able to neutralized with HBsAg. 24 of 30 pieces of HBsAg weak positive samples were judged to be positive, while 6 poeces were negative. The ELISA comfirm method is fully consistent with Roche confirmatory Kit. CONCLUSION: The ELISA confirmatory test for suspicious HBsAg positive samples is a simple, accurate and low cost initial validation method, After further clinical trials, should be widely applied.

[Comparison of two different methods to detect HIV antibodies].

OBJECTIVE: Evaluated the chemiluminescence and enzyme-linked immunosorbent assay (ELISA) to detect HIV antibodies, and compared the results, to provide a reference for the selection and clinical application of HIV screening. METHODS: 3000 cases of our hospital patients were measured by enzyme-linked immunosorbent assay and chemiluminescence immunoassay, using comfirmming experimental results as gold standards. Comparing sensitivity, specificity and other Indicators. RESULTS: In the diagnosis of HIV infection, enzyme-linked immunosorbent assay and chemiluminescence immunoassay had no significant difference. The positive rate of enzyme-linked immunosorbent assay was 0.93%, while the sensitivity and specificity were 89.66%, 99.93%, the positive rate of chemiluminescence immunoassay was 1.03%, while the sensitivity and specificity were 100%, 99.93%, respectively. CONCLUSION: Both methods are suitable for screening of HIV, having high specificity, and chemiluminescence has greater sensitivity than ELISA.

[Analysis for post-transplant changes of peripheral lymphocyte subsets in liver transplant receptors with HBV infection].

AIM: To investigate the characters and changes of peripheral lymphocyte subsets and cytokines in liver transplant receptors with HBV infection in short phases after liver transplantation, and to provide evidences for monitoring post-transplant immune condition of liver transplant receptors. METHODS: Peripheral lymphocyte subsets and cytokine levels in pre- and post- transplant 12 h, 3 d, 10 d, 30 d, 60 d of 20 cases of patients with HBV-associated severe hepatic diseases were investigated and analyzed, and were compared respectively with those of 22 cases of healthy adults as control (HC) with flow cytometry (FCM) and ELISA. RESULTS: The patients' accounts of peripheral lymphocyte subsets before liver transplantation were lower than those of HC significantly, but the accounts decreased significantly after transplantation 12 h. Three days later, the accounts of lymphocyte subsets increased significantly. The percentages of CD3, CD4, CD8 and NK cells got to stable stage from post-transplantation 10 d, and the absolute accounts of post-transplantation 60 d were higher than those of pre-transplantation, but were still lower than those of HC; The IFN-γ and IL-10 levels of post-transplantation 12 h increased several times and decreased after 3 days. The IL-10 levels in post-transplantation 60 d were still higher than those of HC. CONCLUSION: The absolute accounts of peripheral lymphocyte subsets increased to stable levels from post-transplantation 10 d, but were still lower than those of HC; Post-transplant immune condition was to Th2 polarization.

[The verification of a new DNA double-detection diagnostic kit for identifying novel influenza A (H1N1)].

OBJECTIVE: To verify a new kit of "universal and novel influenza A (H1N1) virus nucleic acid double-detection methods (PCR-fluorescence probe)". METHODS: 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. RESULTS: 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. CONCLUSION: 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).