Virological Markers Research Articles

Analysis of Mutations in the'a'Determinant of HBsAg in Chronic Hepatitis B Patients with Coexisting HBsAg and Anti-HBs Positivity

B형 간염바이러스 표면항원에 대한 항체가 형성되면 HBsAg이 사라지고 HBV 감염으로부터 회복되는 것이 보편적이지만, HBsAg이 양성인 만성 간염 환자들 중 지속적으로 anti-HBs가 양성인 경우를 관찰할 수 있다. 이에 저자는 HBsAg과 anti-HBs가 동시에 양성인 만성 B형 간염환자에서 HBsAg S gene의?"a"?결정기 변이 존재 및 임상적 의의를 조사하였다. 53명의 환자 중 47명은 표면항원과 항체가 동시에 양성이었으며, 6명은 표면항원만 양성인 환자였다. 53명의 환자 중 17명(32.1%)이?"a"?결정기 변이가 관찰되었다. 시험군의 4명과 대조군의 1명은?"a"?결정기에 최소한 2개 이상의 변이가 관찰되었다. 가장 빈번한 변이 부위는 aa126, aa131 그리고 aa133였다. 이상의 결과로써 HBsAg과 anti-HBs가 동시에 양성인 B형 간염 환자에서 숙주의 면역체계를 벗어날 수 있는 HBsAg?"a"?결정기 부위에 다수의 변이가 있음을 확인하였다. Serologic markers are used to screen and diagnosis hepatitis B virus(HBV) infection. After an infection with HBV, hepatitis B surface antigen(HBsAg) is the first virologic marker detectable in the serum. If anti-HBs against?"a"?determinant of HBsAg appears, HBsAg will be disappeared and the patients will recover from the HBV infection in most cases. However, the coexistence cases of HBsAg and anti-HBs were frequently observed. The author investigated the clinical significance and presence of mutations of "a"?determinant in the S gene of chronic hepatitis B patients with coexisting HBsAg and anti-HBs. 47 out of the 53 patients were both positive for HBsAg and anti-HBs, 6 were positive for HBsAg only. 17(32.1%) mutations were found within the?"a"?determinant of HBV in the 53 patients. 4 patients from the test group, but only 1 from the control group, presented at least two residue changes in the?"a"?determinant. The most frequent changes in the test group were located at positions aa126, aa131, and aa133. These results demonstrated the presence of mutations at HBsAg?"a"?determinant in chronic hepatitis B patients with concurrent HBsAg and anti-HBs

Nucleic Acid Testing for the Detection of HBV DNA

Dear Editor, We read with interest the article by Arababadi et al. [1], which highlights different and important areas of concern in the management of occult hepatitis B (OBI). There are two relevant issues worthy of discussion. First, the authors described the variable prevalence of OBI in different geographical regions worldwide. The laboratory assay used to detect OBI is an important determinant of its prevalence. Because of the low viremic state of patients, OBI may not be detected even when sensitive polymerase chain reaction (PCR) assays are employed. Results of a recent population study in our locality involving 13,034 blood donors showed the prevalence of OBI to be between 0.11% and 0.13% [2]. We used nucleic acid testing to detect hepatitis B virus (HBV) DNA, which has a detection limit (3.7 IU/mL) lower than that of most available PCR assays. Only studies using sensitive assays should be considered when implementing policies for screening blood donors. Although Arababadi et al. have suggested using an antibody against the hepatitis B core antigen (anti-HBc) for screening blood donors, there is a vast difference in the prevalence of serum anti-HBc positivity and OBI worldwide. For example, the preva-lence of anti-HBc positivity in Korea is 13.5%, whereas the prevalence of OBI in the same region is 0.016% [3]. Hence, in blood donor screening programs, hepatitis B surface antigen (HBsAg) testing followed by nucleic acid testing should be regarded as the most accurate approach for the detection of OBI. Second, the authors have suggested that OBI is a possible risk factor for hepatocellular carcinoma (HCC), but they have also commented that the underlying patho-genesis is unclear. Recently, an Asian study compared the tumorous and adjacent non-tumorous liver tissues from 33 cases of cryptogenic HCC and 28 of HCC with identifiable causes [4]. Both intrahepatic HBV DNA and HBV pregenomic RNA (pgRNA) were detected in 73% of the cases of cryptogenic HCC, albeit at levels lower than those detected in cases of HBV-related HCC. Hence, it is very likely that both persistent viral replication and viral integration play an important role in the pathogenesis of OBI-related HCC. The detection rate of intrahepatic HBV DNA was higher in the non-tumorous tissues than in the tumorous tissues; this finding is consistent with the hypothesis that tumorous tissues lack the optimal environment for HBV replication [5], which may affect the detection of OBI-related HCC. Therefore, in regions with a high prevalence of chronic hepatitis B, OBI should be considered as a possible cause of cryptogenic HCC even when HBV virologic markers are undetectable in tumor histological analysis.

Detection of human T-cell lymphotropic virus type 1 in plasma samples

Human T-cell lymphotropic virus type 1 (HTLV-1) is an RNA virus responsible for diseases such as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T-cell leukemia/lymphoma (ATL). Cell-to-cell contact and Tax-induced clonal expansion of infected cells are the main modes of virus replication, making virus detection during the viremic stage difficult. Consequently, the proviral load is the current virologic marker for disease monitoring, but the mechanisms of progression have not been established yet. Thus, this study investigated the presence of virus in plasma from asymptomatic HTLV-1 carriers and from HAM/TSP patients. Real-time PCR was performed on DNA from 150 plasma samples; 12 (8%) had detectable DNA amplification, including 6 (4%) asymptomatic HTLV-1 carriers and 14 (26%) HAM/TSP patients (p<0.005). Of the 33 samples submitted for nested PCR, six (18%, p=0.02) were positive for HTLV-1 RNA in the plasma. Additionally, 26 plasma samples were treated with DNAse enzyme to eliminate any DNA contamination before RNA extraction. Two of them (8%) showed amplification for HTLV-1 (p=0.5). Therefore, this study described for the first time the detection of free HTLV-1 RNA in plasma from HTLV-1-infected subjects, regardless of their clinical status. Thus, HTLV-1 viral replication does occur in plasma, and other transmission pathways for HTLV-1 should be investigated further.

Kinetics of Viremia and NS1 Antigenemia Are Shaped by Immune Status and Virus Serotype in Adults with Dengue

BackgroundDengue is a major public health problem in tropical and subtropical countries. Exploring the relationships between virological features of infection with patient immune status and outcome may help to identify predictors of disease severity and enable rational therapeutic strategies.MethodsClinical features, antibody responses and virological markers were characterized in Vietnamese adults participating in a randomised controlled treatment trial of chloroquine.ResultsOf the 248 patients with laboratory-confirmed dengue and defined serological and clinical classifications 29 (11.7%) had primary DF, 150 (60.5%) had secondary DF, 4 (1.6%) had primary DHF and 65 (26.2%) had secondary DHF. DENV-1 was the commonest serotype (57.3%), then DENV-2 (20.6%), DENV-3 (15.7%) and DENV-4 (2.8%). DHF was associated with secondary infection (Odds ratio = 3.13, 95% CI 1.04–12.75). DENV-1 infections resulted in significantly higher viremia levels than DENV-2 infections. Early viremia levels were higher in DENV-1 patients with DHF than with DF, even if the peak viremia level was often not observed because it occurred prior to enrolment. Peak viremias were significantly less often observed during secondary infections than primary for all disease severity grades (P = 0.001). The clearance of DENV viremia and NS1 antigenemia occurs earlier and faster in patients with secondary dengue (P<0.0001). The maximum daily rate of viremia clearance was significantly higher in patients with secondary infections than primary (P<0.00001).ConclusionsCollectively, our findings suggest that the early magnitude of viremia is positively associated with disease severity. The clearance of DENV is associated with immune status, and there are serotype dependent differences in infection kinetics. These findings are relevant for the rational design of randomized controlled trials of therapeutic interventions, especially antivirals.

Gestational and Fetal Outcomes in B19 Maternal Infection: a Problem of Diagnosis

Parvovirus B19 infection during pregnancy is a potential hazard to the fetus because of the virus' ability to infect fetal erythroid precursor cells and fetal tissues. Fetal complications range from transitory fetal anemia and nonimmune fetal hydrops to miscarriage and intrauterine fetal death. In the present study, 72 pregnancies complicated by parvovirus B19 infection were followed up: fetal and neonatal specimens were investigated by serological and/or virological assays to detect fetal/congenital infection, and fetuses and neonates were clinically evaluated to monitor pregnancy outcomes following maternal infection. Analysis of serological and virological maternal B19 markers of infection demonstrated that neither B19 IgM nor B19 DNA detected all maternal infections. IgM serology correctly diagnosed 94.1% of the B19 infections, while DNA testing correctly diagnosed 96.3%. The maximum sensitivity was achieved with the combined detection of both parameters. B19 vertical transmission was observed in 39% of the pregnancies, with an overall 10.2% rate of fetal deaths. The highest rates of congenital infections and B19-related fatal outcomes were observed when maternal infections occurred by the gestational week 20. B19 fetal hydrops occurred in 11.9% of the fetuses, and 28.6% resolved the hydrops with a normal neurodevelopment outcome at 1- to 5-year follow-up. In conclusion, maternal screening based on the concurrent analysis of B19 IgM and DNA should be encouraged to reliably diagnose maternal B19 infection and correctly manage pregnancies at risk.

Prevalence and clinical significance of hepatitis D virus co‐infection in patients with chronic hepatitis B in Korea

Hepatitis D virus (HDV) infection can cause severe acute and chronic liver disease in patients infected with hepatitis B virus (HBV). Despite the significant decline in the global HDV infection, it remains a major health concern in some countries. This study aimed to investigate the prevalence and clinical features of HDV co-infection in patients with chronic HBV infection in Korea, where HBV infection is endemic. Nine hundred forty patients [median age, 48 (18-94) years; men, 64.5%] infected chronically with HBV were enrolled consecutively. All patients who were positive for hepatitis B surface antigen (HBsAg) for at least 6 months and were tested for anti-HDV. A portion of the HDV delta antigen was amplified, sequenced, and subjected to molecular and phylogenetic analysis using sera from the patients who were anti-HDV positive. Clinical features and virologic markers were evaluated. Inactive HBsAg carriers, chronic hepatitis B, cirrhosis and hepatocellular carcinoma accounted for 29.5%, 44.7%, 17.9%, and 8.0%, respectively. Only three patients were positive for anti-HDV, corresponding to a 0.32% positive rate. All patients who were positive for anti-HDV were inactive HBsAg carriers. HDV RNA could be amplified by PCR from the sera of two patients. Phylogenetic analysis showed that both carried HDV genotype 1. In conclusion, the prevalence of HDV infection is very low (0.32%) in Korea. All HDVs were genotype 1 and detected in inactive HBsAg carriers. Therefore, HDV co-infection may not have a significant clinical impact in Korean patients with chronic HBV infection.

Could Complement C4 be an Alternative to Biopsy for Chronic Hepatitis B Histopathologic Findings?

Hepatitis B leads to chronic liver disease, cirrhosis, and hepatocellular cancer. Viral markers and other laboratory tests used in diagnosis and follow-up of chronic hepatitis B (CHB) do not correlate well with disease activity and liver histopathology. For this reason, alternative tests that indicate disease activity are needed. We aimed to investigate the utility of serum complement levels for follow-up in patients with CHB with normal and high transaminase levels. One hundred forty-three patients that were evaluated between 2009 and 2010 were included in the study. Hepatitis B early antigen negative CHB cases with high transaminase levels were evaluated as the first group, and cases with normal transaminase level (inactive hepatitis B surface antigen carrier) as the second group, patients with cirrhosis were included as a third group. Age, sex, hepatitis B surface antigen, anti-HBcAg IgM, hepatitis B early antigen, anti-δ, anti-HCV, anti-HIV, serum hepatitis B virus (HBV) DNA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltranspeptidase (GGT), complement C3, and C4 levels of both groups were compared. The relationship between Knodell histologic activity index (HAI) score and fibrosis in liver biopsy specimens and serum complement levels of cases with high transaminase levels were investigated. There were 49 patients with CHB with high transaminase levels; (Female/Male: 22/27). Mean age was 42.3±15.7 y, ALT=104.41±101.74, AST=69.7±65.2, GGT=35.37±20.4, C3 level=104.2±28.8, C4=16.11±4.17, and HBV DNA >2000 IU/mL (>105 copies/mL) in all cases. Remaining 27 patients had cirrhosis. There were 67 patients with CHB with normal transaminase levels (Female/Male: 32/35). Mean age was 39.56±12.9 y, ALT=22.7±5.5, AST=22±5.18, GGT=48.8±60.4, C3=117.85±22.15, and C4=21.44±5.46. Serum complement C4 level in 4 of the CHB cases with normal transaminase levels was low. Serum C3 (P=0.024) and C4 (P=0.001) levels in patients with CHB with high transaminase level were significantly lower. Low serum complement levels were negatively correlated with Knodell-HAI scores in patients with high transaminase levels (r=-0.84; P<0.001). There was no correlation between HAI and HBV DNA, AST, ALT, and GGT. There was no significant correlation between complement C3 and C4 levels and ALT, AST, HBV DNA, and GGT in any of the groups. Child score in patients with cirrhosis negatively correlated with both C3 (P=0.001) and C4 levels (P=0.001). Complement levels in patients with cirrhosis and CHB with high transaminase levels did not significantly differ. Serum complement C4 levels (in contrast to virologic markers and transaminases) significantly correlate with liver biopsy findings and may be a useful indicator of disease activity and/or damage in patients with CHB with high transaminase levels.

Distribution and clinical significance of HBV genotypes in patients with HBV infection in 30 regions of China

To explore the distribution and clinical significance of HBV genotypes in patients with HBV infection in China. Serum samples were collected from 2922 patients with HBV infection. HBV genotyping was performed with type-specific primers polymerase chain reaction, and the virological and biochemical markers were detected, which differences in the genotypes distribution between various regions and liver function and virological markers between various HBV genotyping were analyzed. The genotype B, C, B + C, D of 2922 patients with HBV infection accounted for 15.9%, 83.5%, 0.41%, 0.21% respectively. In Northern China, genotype C was most prevalent, accounting for 90% of all cases, while it was less common in Southern China; genotype C was present in Zhejiang and Jiangsu provinces, but genotype B was comparatively more common in Guangdong, Hunan, Hubei, and Jiangxi provinces. B, C genotype HBV infection patients in the sex difference was not statistically significant; B genotypes compared with C genotype HBV infection patients, the average age of is less (P < 0.001); HBeAg positive rate of C genotype HBV infection patients are higher than that of B genotype (P = 0.023); Viral load of genotype C HBV infection patients is higher than that of genotype B (P = 0.038); Cholinesterase and Albumin levels of genotype C HBV infection patients are lower than that of genotype B (P values were 0.016, <0.001). There were HBV genotype B, C, B + C and D in Chinese patients with HBV infection, with genotype B and C being the major ones. Mainly in northern regions of genotype C, C genotype significantly reduced the southern region, some of the southern region dominated by B genotype. Genotype C HBV infection patients are older, and their HBeAg-positive rate is higher, and their liver damage is more severe, but their viral load is less.

Is interleukin-8 an additional to histopathological changes diagnostic marker in HCV-infected patients with cryoglobulinemia?

The aim of this study was to analyze histopathological changes in the liver and serum inflammatory cytokine level in hepatitis C virus (HCV)-infected patients with and without cryoglobulinemia. The study group consisted of 34 patients with chronic hepatitis C, confirmed by serological and virological markers. Ten out of 34 patients had cryoglobulinemia. The control group consisted of 21 healthy persons. Liver biopsy specimens of HCV-infected patients were evaluated by light microscopy using the grade and the stage according to Batts and Ludwig classification. The quantitative measurements of IL-1β, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor in sera were performed by flow cytometry. The mean age of HCV-infected patients with cryoglobulinemia was higher than age of HCV-infected patients without cryoglobulinemia. Microscopic examination of liver biopsy specimens revealed necroinflammatory activity slightly more prominent in patients with cryoglobulinemia. The most prominent inflammatory changes connected with abundant lymphoid aggregates in most of the examined portal tracts and piecemeal necroses were diagnosed in patients with several extrahepatic manifestations, such as cutaneous manifestations, nephrotic syndrome, polyneuropathy, and arthropathy. Liver fibrosis was similar in patients with and without cryoglobulinemia. The serum levels of all proinflammatory cytokines, especially IL-8, were significantly higher in the patients with cryoglobulinemia in comparison with the patients without cryoglobulinemia and healthy persons. All microscopic features did not correlate with the level of any investigated proinflammatory cytokines.

Infección por virus de hepatitis B (VHB) en inmigrantes subsaharianos en Almería

There is scant information available in Spain regarding virological markers and clinical status in Sub-Saharan patients infected with HVB. A cross-sectional and retrospective study of virological markers and clinical status of HBV infection in 510 adult patients from Sub-Saharan Africa, not co-infected with HIV, most of them from West Africa countries. A total of 90.8% of patients had markers of HBV infection and 137 (26.9%) were HBsAg positive. Among patients with HBsAg positive, 55.9% were chronic inactive carriers. The predominant genotype was E. The study shows a high prevalence of both markers of HBV infection and of chronic hepatitis B in immigrants from Sub-Saharan Africa.

Anti-Hdv Immunoglobulin M Testing in Hepatitis Delta Revisited: Correlations with Disease Activity and Response to Pegylated Interferon-α2A Treatment

The role of anti-HDV immunoglobulin M (IgM) testing in patients receiving pegylated interferon-α therapy for hepatitis delta is unknown. We performed anti-HDV IgM testing in a well defined cohort of HDV-infected patients who were treated with pegylated interferon-α2a plus adefovir, or either drug alone. Sera from 33 HDV-RNA-positive patients from the international HIDIT-1 trial were available for anti-HDV IgM testing (ETI-DELTA-IGMK-2 assay, DiaSorin, Saluggia, Italy) before therapy, at treatment weeks 24 and 48, and at 24 weeks after the end of treatment. Anti-HDV IgM tested positive in 31 out of the 33 patients (94%) prior to treatment. HDV IgM levels correlated with histological inflammatory activity (r=0.51, P<0.01) and were higher in patients with alanine aminotransferase and γ-glutamyl transpeptidase levels above the median (P<0.05). Quantitative anti-HDV IgM values declined in patients responding to antiviral therapy, however anti-HDV IgM remained positive after treatment in the majority of virological responders. We suggest that anti-HDV IgM testing might give additional useful information to determine disease activity in hepatitis delta and to predict treatment response to antiviral therapy with type I interferons. However, determination of anti-HDV IgM can not substitute HDV RNA testing, which remains the primary virological marker for response to therapy.

KSHV DNA viremia correlates with low CD4+ cell count in Italian Males at the time of diagnosis of HIV infection

To evaluate the relevance and the virological and immunological markers of Kaposi sarcoma herpesvirus 8 (KSHV) viremia in Italian male patients at the time of diagnosis of infection with HIV-1, 481 men infected with HIV were recruited consecutively. The presence of KSHV DNA was evaluated in peripheral blood mononuclear cells (PBMCs) and in plasma and correlated with demographic and viro-immunological parameters. Seventy-four patients had KSHV DNA detected in PBMCs. By univariate analysis, the presence of KSHV DNA was associated significantly with unprotected homosexual relationships (P=0.003) and it was significantly higher in patients with CD4+ cell <350 (P=0.025). By multivariate analysis, homosexual relationships were associated independently with KSHV DNA in PBMCs (OR: 3.25; 95% CI: 1.1-9.7; P=0.035). Among the 74 patients with KSHV DNA detected in PBMCs, plasma samples from 60 were analyzed and 33 were positive for KSHV DNA. The CD4+ cell counts and percentages were significantly lower in patients with KSHV DNA in both PBMCs and plasma as compared to patients with only KSHV DNA in PBMCs (P=0.006 and P=0.019, respectively). Among the patients with KSHV DNA detected in PBMCs, all 13 patients with CD4+ cells count <200 had detectable levels of KSHV in their plasma. By multivariate analysis adjusted for the epidemiologic and virological parameters, low CD4+ cell count was the only independent variable associated with the presence of KSHV DNA in plasma (OR, 0.001; 95% CI: <0.001-0.001; P=0.03). In HIV-positive antiretroviral therapy-naïve males, KSHV active replication as detected by KSHV DNA in plasma was associated significantly with low CD4+ cell count.

Treatment of Genotype 4 Hepatitis C Recurring After Liver Transplantation Using a Combination of Pegylated Interferon Alfa-2a and Ribavirin

Hepatitis C virus (HCV) recurrence after liver transplantation (LT) is universal and tends to be more aggressive. Data on post-transplant HCV genotype 4 treatment is scarce. The aim of this study is to assess the safety and efficacy of pegylated interferon alpha-2a (PEG-IFN) in combination with ribavirin in the treatment of recurrent HCV genotype 4 after LT. Twenty-five patients infected with HCV genotype 4 were treated with PEG-IFN alpha-2a at a dose of 180 μg/week in addition to 800 mg/day of ribavirin (the dose was adjusted within the tolerated range of 400-1,200 mg). Pretreatment liver biopsies were obtained from all patients. Biochemical and virological markers were assessed before, during, and after treatment. Twenty-two patients (88%) achieved an early virological response (EVR) (12 patients tested negative for HCV-RNA). Fifteen (60%) and 14 patients (56%) achieved an end of treatment virological response (ETVR) and a sustained virological response (SVR), respectively. Five patients had advanced pretreatment liver fibrosis. Pretreatment ALT was elevated in 24 patients (96%). The most common adverse effects were flu-like symptoms and cytopenia. Eighteen patients (72%) required erythropoietin alpha and/or granulocyte-colony stimulating factor as a supportive measure. One patient developed severe rejection complicated by sepsis, renal failure, and death. Other adverse effects included depression, mild rejection, impotence, itching, and vitiligo. Post-transplant treatment with pegylated interferon alpha-2a and ribavirin achieved SVR in 56% of liver transplant recipients with chronic HCV genotype 4 infection. The combination was relatively safe and exhibited a low rate of treatment withdrawal.

Prognostic markers of symptomatic congenital human cytomegalovirus infection in fetal blood

To identify fetal cord blood prognostic markers of symptomatic congenital human cytomegalovirus infection (HCMV). Retrospective observational study. Fetal medicine unit in Milan and Medical virology unit in Pavia, Italy. HCMV-infected and -uninfected fetuses of mothers with primary HCMV infection during the period 1995-2009. Overall, 94 blood samples from as many fetuses of 93 pregnant women experiencing primary HCMV infection were examined for multiple immunological, haematological and biochemical markers as well as virological markers. Congenital HCMV infection was diagnosed by detection of virus in amniotic fluid, and symptomatic/asymptomatic infections were determined by ultrasound scans, nuclear magnetic resonance imaging, histopathology or clinical examination at birth. Blood sample markers were retrospectively compared in symptomatic and asymptomatic fetuses with congenital infection. A statistical analysis was performed to determine the value of each parameter in predicting outcome. Univariate analysis showed that most nonviral and viral markers were significantly different in symptomatic (n = 16) compared with asymptomatic (n = 31) fetuses. Receiver operator characteristics analysis indicated that, with reference to an established cutoff for each marker, the best nonviral factors for differentiation of symptomatic from asymptomatic congenital infection were β(2) -microglobulin and platelet count, and the best virological markers were immunoglobulin M antibody and DNAaemia. β(2) -Microglobulin alone or the combination of these four markers reached the optimal diagnostic efficacy. The determination of multiple markers in fetal blood, following virus detection in amniotic fluid samples, is predictive of perinatal outcome in fetuses with HCMV infection.

Molecular epidemiology of chronic hepatitis B virus infection in Greece

Virological data on chronic hepatitis B virus (HBV) infection in Greece are limited. HBV genotypes, surface antigen (HBsAg) subtypes, and HBsAg "a" determinant mutations among patients infected chronically with HBV, were investigated. Serum samples from 135 HBsAg positive patients were tested. Serologic (HBsAg, anti-HBs, HBeAg, and anti-HBe), virologic (HBV-DNA quantitation) and biochemical markers (serum alanine aminotransferase/ALT and aspartate aminotransferase/AST) were analyzed. HBV genotypes and HBsAg subtypes were determined by partial sequencing of the S gene. Genotyping was performed by using the National Center for Biotechnology Information online Genotyping tool and phylogenetic analysis. Nucleotide sequences were aligned pair wise with ClustalW and phylogenetic trees were constructed by the neighbor-joining method. Sequences were also used to predict HBV HBsAg subtypes. In six patients (4%), simultaneous presence of HBsAg and anti-HBs was determined, whereas 47 patients (35%) were HBeAg positive, 84 (62.5%) were anti-HBe positive, and four patients (3%) were characterized by the simultaneous presence of HBeAg and anti-HBe. Mean ALT was 238 IU/L (standard deviation = 576.84), and HBV-DNA levels ranged from 1.02 × 10(5) to 2.2 × 10(7) IU/ml. Genotype D was predominant (98%), with viral groups D/ayw2 (73%) and D/ayw3 (27%). Group A/adw accounted for 1% of cases. Genotypes B and C were found exclusively in the Chinese immigrants (1%). Single or multiple point mutations were found in 35 cases (26%). Some of the most common mutations occurred at amino acid positions 129, 133, 134, 144, 145, including the "vaccine escape" mutation G145R. Mutations analysis revealed that amino acid substitutions did not affect detection by commercial immunoassays.

Correlation of Immune Activation during Late Pregnancy and Early Postpartum with Increases in Plasma HIV RNA, CD4/CD8 T Cells, and Serum Activation Markers

A previously observed rise in the plasma viral load postpartum in both treated and untreated HIV-positive women remains unexplained. Virological and immunological markers were evaluated in HIV-negative controls and HIV-positive pregnant women with and without antiretroviral treatment. Plasma HIV RNA, CD4/CD8 T cells, and serum activation markers were sequentially measured during the third trimester, at delivery, and 2 to 8 weeks postpartum in a cohort of HIV-positive pregnant women (n = 96) enrolled in a maternal-fetal HIV transmission study and a control group of HIV-negative pregnant women (n = 28). Mean plasma HIV RNA (P = 0.003) increased from delivery to postpartum, and mean CD4 T cells (P = 0.002) and serum β2-microglobulin (P < 0.0001) increased from the third trimester through postpartum among the HIV-positive women. Mean CD8 T cells increased from the third trimester through postpartum in women receiving zidovudine (ZDV) and in those not treated (P < 0.05) but remained stable in those on highly active antiretroviral therapy (HAART) and the HIV-negative controls. Increases in serum β2-microglobulin were correlated with increases in HIV RNA (P = 0.01). HIV-positive pregnant women showed postpartum increases in plasma HIV RNA, CD4 T cells, and serum β2-microglobulin regardless of the treatment regimen. The rise in CD4 T cells and β2-microglobulin was also observed in HIV-negative pregnant women, suggesting hormonal changes and/or labor-induced cytokines may contribute to immune activation. Immune activation correlated with increased plasma HIV RNA in postpartum women despite treatment, although HAART appeared to blunt the effect. The observed rise in plasma HIV RNA postpartum, which correlated with markers of immune activation, may have implications for enhanced transmission to infants through early breast-feeding and to sexual partners.

Childhood myocarditis and parvovirus B19 genotypes

Background Human parvovirus B19 (PVB19) infection is occasionally associated with acute myocarditis. Three cases of children with PVB19 virus-associated myocarditis occurred in a very short period and the same geographical region. Objective To elucidate if virological factors could be responsible for determining the course of infection, a molecular epidemiologic investigation was performed. Study Design The diagnosis of myocarditis was established by histology or echocardiography. In the three cases, the PVB19 DNA was detected in different samples. Eight different regions were amplified by PCR using a high fidelity Taq polymerase and sequenced on both strands. Phylogenetic analyses were performed. First, the genotypes of the PVB19 strains were determined, then the intra-patient viral variability was analysed by sequencing PVB19 detected in different specimens sampled from the same patient at the same moment. Results Nearly complete sequences of the PVB19 virus (4265nt) were obtained from different samples in the three patients. The phylogenetic analyses showed that PVB19 strains identified clustered with genotype 1a PVB19 strains referenced in GenBank. When compared to the referenced strain NC_000883, the number of substitutions (transitions and transversions) were as follows: 58 for Caen.FRA/19.09, 74 for Caen.FRA/21.09 and 60 for Caen.FRA/24.09. The strains isolated from the same patient showed 100% of similarity. Conclusions Viral myocarditis is a frequently unrecognized cause of post-inflammatory cardiomyopathy. The detailed molecular analyses do not give rise to virological markers associated with myocarditis in these children.

27 First case of HIV window donation in Switzerland after 8 years of universal HIV NAT screening

Transfusion-associated HIV transmission is a rare but dreaded event. About 350’000 donations are annually screened for human-immunodeficiency virus (HIV) in Switzerland. Anti-HIV antibody screening by ELISA testing may fail in early infection because of a diagnostic window-phase of about 21 days. Therefore, blood donations in Switzerland are screened for HIV by nucleic acid test (NAT) since 2002. The diagnostic window phase for HIV will be diminished by NATscreening. At an average the remaining time period accounts for 5 to 6 days. The adjacent figure shows the kinetics of virological markers and the host immune response in the early phase of HIV infection. The p24-antigen will be detected about 14 days after infection, as can be inferred from the graph.

Estimation of the Distribution of Infection Times Using Longitudinal Serological Markers of HIV: Implications for the Estimation of HIV Incidence

In the last decade, interest has been focused on human immunodeficiency virus (HIV) antibody assays and testing strategies that could distinguish recent infections from established infection in a single serum sample. Incidence estimates are obtained by using the relationship between prevalence, incidence, and duration of recent infection (window period). However, recent works demonstrated limitations of this approach due to the use of an estimated mean "window period." We propose an alternative approach that consists in estimating the distribution of infection times based on serological marker values at the moment when the infection is first discovered. We propose a model based on the repeated measurements of virological markers of seroconversion for the marker trajectory. The parameters of the model are estimated using data from a cohort of HIV-infected patients enrolled during primary infection. This model can be used for estimating the distribution of infection times for newly HIV diagnosed subjects reported in a HIV surveillance system. An approach is proposed for estimating HIV incidence from these results.

Kinetics of dengue virus NS1 protein in dengue 4-confirmed adult patients

In this work, the presence of NS1 protein as a possible early marker of dengue infection was studied in serum samples from confirmed adult patients with a primary and secondary dengue 4 infection. A total of 209 serum samples collected from day 2 up to day 7 of fever onset from 71 patients were tested by Platelia NS1 antigen capture ELISA kit (BioRad, Marnes-la-Coquette, France), and the results were compared with those obtained by capture antidengue virus IgM (MAC)-ELISA and ELISA inhibition method tests. The 83.3% of primary cases and 96.4% of secondary cases were NS1 positive. The kinetics of NS1 protein showed the highest values in optical density mean ratio or in percentage of positives between days 2 and 4. The results obtained in this study show the utility of the NS1 protein as a virologic early marker of dengue infection. Prospective studies should be carried out to confirm its utility as a prognostic marker of severe illness.