China ; DNA, Viral ; blood ; Hepacivirus ; classification ; Hepatitis C ; diagnosis ; etiology ; therapy ; Humans ; RNA, Viral ; analysis

OBJECTIVETo compare the detection rates of Epstein-Barr virus (EBV) DNA in the serum/plasma between apparently healthy adults (AHAs) and nasopharyngeal carcinoma (NPC) patients in attempt to evaluate the efficiency of EBV DNA assay for serodiagnosis of NPC.

METHODSThe plasma and serum were obtained from 58 AHAs and 66 untreated NPC patients. EBV DNA W-fragment was detected using nested ploymerase chain reaction (PCR). Immunoenzymatic assay for titration of IgA-VCA was also adopted.

RESULTSEBV DNA detection rate (84.85%) in the plasma/serum of 66 NPC patients was significantly higher than that (10.34%) in 58 AHAs. The sensitivity of plasma/serum EBV DNA assay (0.8485) was higher than that (0.8030) of titrating IgA-VCA (positive criterion >/= 1:40) though the specificities of these two tests were the same (0.8966). The correct rate, predictive value of a positive test, and Odds ratio of dual positivity (0.8387, 0.9792 and 141.0, respectively) were higher than those of single positivity either to plasma/serum EBV assay (0.5242, 0.7333 and 1.1423, respectively) or to IgA-VCA >/= 1:40 test (0.4839, 0.5385 and 1.0480, respectively).

CONCLUSIONThe EBV DNA detection in the plasma/serum using nested PCR may be a useful indicator for serodiagnosis of NPC.

Antigens, Viral ; blood ; DNA, Viral ; blood ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunoglobulin A ; blood ; Nasopharyngeal Neoplasms ; diagnosis ; virology ; Serologic Tests

OBJECTIVETo use dried blood spot (DBS) in studying the sequence and subtype analysis of HIV-1 genome.

METHODS2 ml whole blood containing EDTA anticoagulant from 20 HIV-1 infected patients were collected, then 80 microl blood was used to propare DBS. QIAamp Blood Mini kit and 10% Chelex100 resin extracted DNA genome from whole blood and DBS as well as nested PCR amplified specifically HIV-1 Gp41 region from the two kinds of DNA extraction. Software MAGE 3.0 was used to study the sequence and subtype of the PCR products.

RESULTSEligible 18 paired samples were analysed to show that 16 of them belonged to C54A, 97CNGX-7F, 98CN006 subtypes. The other two samples might belong to B. CN._. RL42 and B. US. 90WEAU160.

CONCLUSIONData showed that there were parallel results between the whole blood and DBS samples including subtype analysis, position of mutation and types of amino acid sequencing. Since DBS itself facilitated the collection, transportation and storage, it could be used as a measure to collect blood sample in resource limited area and to develop molecular epidemiologic research as well as early diagnosis on infant exposed to HIV.

DNA, Viral ; blood ; HIV Infections ; blood ; genetics ; HIV-1 ; genetics ; Humans ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; methods

OBJECTIVETo explore the changes in HBsAg titer and HBV DNA load and their correlation in patients with chronic hepatitis B (CHB) and HBV-related liver cirrhosis (HBV-LC).

METHODSForty-six patients with mild to moderate CHB (CHB-LM), 24 patients with severe CHB (CHB-S), and 28 patients with HBV-LC at admission, and 51 patients with HBV-LC at 4.08 ± 3.06 months during antiviral treatment were tested for serum HBsAg titer and HBV DNA load using Abbott chemiluminescence and fluorescence quantitative PCR, respectively.

RESULTSThe serum HBsAg titer and HBV DNA load gradually decreased with increased disease severity (from CHB-LM, CHB-S to HBV-LC; χ(2)=12.537 and 8.381, respectively, P<0.05). HBsAg titer and HBV DNA load were significantly higher in CHB-LM and CHB-S groups than in HBV-LC group (P<0.05), but comparable between CHB-LM and CHB-S groups (Z=-0.649 and 0.032, respectively, P>0.05). Among HBeAg-positive patients, HBsAg titer and HBV DNA load tended to decrease with increased disease severity (from CHB-LM, CHB-S to HBV-LC; χ(2)=6.146, P=0.046 and χ(2)=1.017, P>0.05; respectively), and CHB-LM group had significantly higher HBsAg titer than HBV-LC group (Z=-2.247, P=0.025). Among the HBeAg-negative patients, serum HBsAg and HBV DNA load gradually declined with the disease severity (χ(2)=8.660 and 13.581, respectively, P<0.05), and were obviously higher in CHB-LM and CHB-S groups than in HBV-LC group (P<0.05). Positive correlations were found between serum HBsAg and HBV DNA levels in CHB-LM (r=0.389, P=0.009) and HBV-LC groups (r=0.431, P=0.022), but not in CHB-S group (r=0.348, P=0.104). After antiviral therapy, the serum HBsAg titer was slightly decreased (Z=-1.050, P=0.294) while HBV DNA load markedly reduced (Z=-5.415, P<0.001), showing no correlation between them (r=0.241, P=0.111) or between the measurements before and after treatment (r=0.257, P=0.085).

CONCLUSIONSerum HBsAg titer and HBV DNA load decreases progressively from CHB-LM to CHB-S and HBV-LC in both HBeAg- positive and -negative patients. The serum HBsAg titer is positively correlated with HBV DNA load, but their levels are not consistently parallel.

Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; blood ; Humans ; Liver Cirrhosis ; blood ; virology ; Viral Load

OBJECTIVETo 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.

METHODSIntrahepatic 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.

RESULTSIntrahepatic 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).

CONCLUSIONIn 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.

DNA, Circular ; analysis ; DNA, Viral ; analysis ; Female ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Liver ; virology ; Male ; Viral Load

OBJECTIVETo assess the value of quantitative analysis of hepatitis B surface antigen (HBsAg) levels in the diagnosis and therapeutic evaluations in patients with chronic hepatitis B (CHB).

METHODSAccording to the staging criteria defined by the American Association of Liver Diseases, 96 patients with CHB admitted in Zhujiang Hospital were classified in immune-tolerant (IT), HBeAg-positive hepatitis (EPH), inactive carrier (IC) and HBeAg-negative hepatitis (ENH) phases. Serum HBsAg, HBV-DNA and ALT levels were quantified and their correlations were evaluated in each phase of infection.

RESULTSThe mean HBsAg titers (measured in log10U/L) differed significantly between the phases of CHB (4.12 in IT, 4.02 in EPH, 2.85 in EPH, and 3.29 in ENH). The correlation coefficient of HBsAg with HBV-DNA was 0.6828 in IT, 0.5759 in EPH, 0.3280 in IC, and 0.1083 in ENH. Serum HBsAg titers were significantly higher in HBeAg-positive patients than in HBeAg-negative patients. No correlation was found between HBsAg level and ALT in each phase of CHB.

CONCLUSIONThe median baseline serum HBsAg levels vary between different phases of CHB in Guangzhou, suggesting the value of HBsAg in accurate classification of hepatitis B patients and evaluation of the therapeutic effect and outcomes of the patients.

DNA, Viral ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; Hepatitis B, Chronic ; blood ; Humans ; Serologic Tests

OBJECTIVE: To determine the expression profile of microRNA (miRNA) in peripheral blood mononuclear cells (PBMC) and immune factors in pregnant women with hepatitis B virus (HBV) infection. METHODS: A total of 182 pregnant women infected with HBV were randomly selected, with 40 healthy pregnant women and 35 non-pregnant women as controls. High-throughput sequencing was used to detect RNA in the PBMC of all subjects. Indirect ELISA method was used to determine the changes of cytokines in peripheral blood samples. RESULTS: Compared with the control group, 18 differentially expressed miRNA were identified in those with HBV infection (P< 0.01). Among these, miR-3607-3p, miR-20a, miR-1296, miR-153-1 and miR-X4 may directly regulate the transcriptional level of target genes including IL-10, IL-18, IL-16, MCP-1, NUP50 and CCR1. Meanwhile, peripheral blood cytokines IL-10, IL-18, IL-16 and MCP-1 were significantly increased in those with HBV infection (P<0.01), with the expression level of IL-16 and MCP-1 being strongly correlated with the viral load. CONCLUSION The expression profiles of miRNA in PBMC and cytokines in peripheral blood can change significantly during pregnancy, both may be involved in the immune response to HBV infection.
Cytokines ; blood ; DNA, Viral ; Female ; Hepatitis B ; blood ; Hepatitis B virus ; Humans ; Leukocytes, Mononuclear ; metabolism ; MicroRNAs ; blood ; Pregnancy

OBJECTIVE: To analyze the relationship between serum HBsAg concentration and HBV replication level in hepatitis B patients with positive serum HBsAg and HBeAg, and to explore the possibility of using serum HBsAg concentration as a marker of HBV replication level in hepatitis B patients with positive serum HBeAg. METHODS: HBV DNA level and serum HBeAg, HBsAg concentration of 296 patients with positive serum HBsAg and HBeAg were quantitatively detected by real-time fluorescence quantitative PCR (FQ-PCR) and time-resolved fluoroimmunoassay (TRIFA) respectively. HBsAg concentrations were compared among patients with different HBV DNA levels, and HBV DNA levels were compared among patients with different HBsAg concentrations. The correlation between serum HBsAg concentration and DNA replication level were analyzed. The positive, negative predictive values and coincidence rates were speculated by various HBsAg concentrations. RESULTS: If HBV DNA positive was defined as HBV DNA levels no less than 10(5) copy/mL, then 228(77.03%) patients were classified as HBV DNA positive. HBsAg concentration was positively correlated with HBV DNA replication level, but among groups with various DNA replication levels, HBsAg concentration showed no significant statistical difference (P>0.05). If the patients were divided into 2 groups, HBsAg concentration (180 microg/L) was served as the cutoff level, the DNA positive rate of the group with HBsAg concentration no less than 180 microg/L was significantly higher than that with HBsAg concentration less than 180 microg/L (chi(2)=3.998, P<0.05). DNA positive rates and average DNA levels showed no significant statistical differences between the 2 groups, if HBsAg concentrations other than 180 microg/L were used as the cutoff level. Positive predictive values, negative predictive values and the coincidence rates speculated by various HBsAg concentrations as cutoff values did not show any significant statistical difference in estimating HBV replication levels. CONCLUSION To some extent, serum HBsAg concentration is related to HBV DNA replication level in hepatitis B patients with positive serum HBsAg and HBeAg, but it is not feasible to use HBsAg concentration to monitor their HBV replication levels.
DNA, Viral ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; virology ; Humans ; Virus Replication

OBJECTIVEHepatitis B virus covalently closed circular DNA in the serum of patients with hepatitis B, peripheral mononuclear cells (PBMC) and liver tissue distribution.

METHODSSerum HBV DNA > 10(5) copies/ml 50 cases of hepatitis B patients, serum HBV DNA < 10(5) copies/ml 30 cases of hepatitis B patients, patients with fatty liver (hepatitis B) 20 cases, using real-time quantitative polymerase chain reaction for detection of serum, and liver tissue in PBMC HBVcccDNA the existence.

RESULTSOf 50 serum HBV DNA > 10(5) copies/ml cccDNA the serum specimens were 28 cases, 56% detection rate, 29 cases were PBMC HBVcccDNA, 58% detection rate, liver tissue HBVcccDNA were 44 cases, 88% detection rate, serum, the PBMC were detected in liver tissue were significant differences in P < 0.005, compared serum PBMC were no significant differences in P > 0.005. 30 serum HBV DNA < 10(5) copies/ml the serum specimens, PBMC cccDNA detected two cases were 6.67% detection rate, liver tissue cccDNA were six cases 20% detection rate, serum, PBMC, were among the liver tissue was not significantly different P > 0.005. 20 in the serum of patients with fatty liver, liver tissue in PBMCwere not detected HBVcccDNA.

CONCLUSIONHBVcccDNA mainly in the liver of patients with hepatitis B, hepatitis B patients and also cccDNA PBMC in the presence of liver tissue but a few of many.

Adult ; DNA, Circular ; blood ; DNA, Viral ; blood ; Fatty Liver ; blood ; virology ; Female ; Hepatitis B ; blood ; virology ; Hepatitis B virus ; genetics ; physiology ; Humans ; Liver ; virology ; Male

OBJECTIVETo investigate the relationship between HBV DNA and hepatitis B virus large envelope protein (HBV-LP) in patients with chronic hepatitis B.

METHODSSerum HBV DNA was detected by RT-PCR and the HBV-LP was detected by enzyme linked immunosorbent assay in 320 serum samples collected from patients with chronic hepatitis B.

RESULTSThere were no significant difference between positive rate of HBV-LP and that of HBV DNA in different HBeAg patterns (P>0.05). Serum HBV-LP levels were closely correlated with HBV DNA copies (r=0.949).

CONCLUSIONSerum HBV-LP is a reliable serological marker that can reflect the replication of HBV.

Adolescent ; Adult ; Aged ; DNA, Viral ; blood ; Female ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Male ; Middle Aged ; Viral Envelope Proteins ; blood ; Virus Replication ; Young Adult