Virus DNA Assays Research Articles

Comparison between Quantitative Hepatitis B DNA and HBeAg Positivity to Detect Active Viral Replication among Patients with Hepatitis B infection in a Tertiary Care Hospital

To study the correlation between quantitative Hepatitis B DNA and HBeAg positivity among HBsAg positive patients in a tertiary care hospital. To detect HBsAg, HBeAg levels by CMIA (Chemiluminescent microparticle immunoassay). To perform quantitative Hepatitis B virus DNA assay using Real Time Polymerase Chain Reaction. The study took place between July and December 2022 for 6 months. Sixty four patients were included in the study for whom HBsAg was positive by CMIA and for whom HBeAg (detected by CMIA) and Hepatitis B Virus DNA by quantitative Real Time PCR was performed. Of the 64 HBsAg reactive patients, the ‘e’ antigen of Hepatitis B virus was positive in six patients. In all the six HBeAg positive patients, HBV DNA was detected with a range of 78-10,288 IU/ml with 66% having levels more than 2000 IU/ml and requiring treatment. Among the 58 patients who were negative for the ‘e’ antigen of Hepatitis B virus, the Hepatitis B Viral DNA was detected in 27 patients with a range of 10-1,76,000 IU/ml, with 21% of patients having levels more than 2000 IU/ml. HBeAg is a good serological test to detect viral replication. However, it alone might not be sufficient as quantitative HBV DNA levels are more accurate and it does identify more patients who may require treatment.

Evaluation of Hepatitis B viral Serological Patterns and HBV DNA levels in Hepatitis B subjects at Nauth Nnewi, Southeastern Nigeria

Nigeria is one of the countries with the highest incidence of Hepatitis B Virus (HBV) infection worldwide. The inaccessibility and unaffordability of HBV DNA quantification (viral load) assay which is the key laboratory test for therapy initiation, and monitoring is a challenge to HBV management. This study aimed at evaluating the HBV DNA viral load differences across the serological markers of HBV in order to develop a more cost-effective diagnostic algorithm for Hepatitis B management. Cross sectional study design was used with a total of 264 subjects comprising of 88 HBsAg seropositive treatment naïve subjects, 88 HBsAg seropositive subjects on antiviral therapy as case subjects and 88 age-matched apparently healthy HBsAg seronegative individuals were recruited as control subjects. Hepatitis B Virus DNA assay was performed using real time PCR technique, Hepatitis B core Antibody Immunoglobulin M and Hepatitis D Virus Immunoglobulin G assay. Immunochromatography was used for HBV Panel, Hepatitis C Virus assay and Human Immunodeficiency Virus testing. The non-treatment group has higher viral load (M=805.50 IU/ml) compared with treatment group (M = 65.50 IU/ml) (p<0.001). There was a significant difference in the HBV DNA levels of the four serological patterns observed in the study (P< 0.001). Among the four serological patterns observed, the second pattern with positive surface antibody, positive envelope antigen and negative envelope antibody showed highest viral load (M= 46850189.50 IU/ml) followed by the third pattern with negative surface antibody, negative envelope antigen, and negative envelope antibody (M=46555 IU/ml). The first pattern which has negative surface antibody, negative envelope antigen and positive envelope antibody has the lowest viral load (M=21.00 IU/ml) followed by the fourth pattern with positive surface antibody, negative envelope antigen and positive envelope antibody (M=493.00 IU/ml). This study showed that HBV serological markers can predict viral load and should be used as its alternative in resource poor settings.

Evaluation of HBsAg Quantification as Surrogate to HBV DNA Viral Load in Hepatitis B Infected Patients in Anambra State, Nigeria

Hepatitis B is an infectious disease of great public health importance. Nigeria is one of the countries with the highest incidence of Hepatitis B Virus (HBV) infection worldwide. However, the accessibility and affordability of HBV DNA quantification (viral load) assay is the key laboratory test for therapy initiation, and monitoring is a challenge to HBV management. This study aimed at determining the relationship between HBV DNA quantification and routine haemato-serological parameters in order to develop a more cost-effective diagnostic algorithm for Hepatitis B management. Cross sectional study design was used with a total of 264 subjects comprising of 88 HBsAg seropositive treatment na&#239ve subjects, 88 HBsAg seropositive subjects on antiviral therapy as case subjects and 88 age-matched apparently healthy HBsAg seronegative individuals were recruited as control subjects. Hepatitis B Virus DNA assay was performed using real time PCR technique while ELISA technique was used for Hepatitis B surface antigen quantification. HBsAg quantification showed strong positive correlation with HBV DNA viral load both in treatment and non-treatment groups (r = 0.673; p

Clinical value of a plasma Epstein-Barr virus DNA assay in the diagnosis of recurrent or metastatic nasopharyngeal carcinoma: a meta-analysis.

Background: To evaluate the diagnostic value of Epstein–Barr virus (EBV) DNA in nasopharyngeal carcinoma (NPC) patients with locoregional or distant recurrence.Methods: Articles related to the diagnosis of recurrent or metastatic NPC by the detection of EBV DNA in plasma or serum were retrieved from different databases. Sensitivity, specificity, summary receiver operating characteristic (SROC) curves, and likelihood ratios were pooled to assess the diagnostic value of individual diagnostic tests.Results: This meta-analysis pooled 25 eligible studies including 2496 patients with NPC. The sensitivity, specificity, positive likelihood ratio (+LR), and negative likelihood ratio (−LR) of EBV DNA in the diagnosis of NPC were 0.858 (95% confidence interval (CI): 0.801–0.901), 0.890 (95% CI: 0.866–0.909), 7.782 (95% CI: 6.423–9.429) and 0.159 (95% CI: 0.112–0.226), respectively. The diagnostic odds ratio (DOR) was 48.865 (95% CI: 31.903–74.845). The SROC for EBV DNA detection was 0.93 (95% CI: 0.90–0.95).Conclusion: The detection of EBV DNA for the diagnosis of recurrent or metastatic NPC has good sensitivity and specificity and might be helpful in monitoring recurrent or metastatic NPC.

Commutability and concordance of four hepatitis B virus DNA assays in an international multicenter study.

BackgroundHBV DNA is the most important molecular marker in hepatitis B, used to determine treatment indication and monitoring. Most patients require lifelong hepatitis B virus (HBV) management, thus viral load (VL) monitoring may be performed at different laboratories, with different HBV assays, which may result in different VL results. This multicenter study compares the commutability and concordance of results from four different HBV DNA assays: CAP/CTM HBVv2, HPS/CTM HBVv2 and the new cobas 6800/8800 HBV and cobas 4800 HBV assays.MethodsAcross all four assays, HBV limit of detection (LoD) and linearity at lower concentrations were assessed using panels traceable to the World Health Organization international standard, and concordance was investigated at the important medical decision cutoffs 2000 and 20,000 IU/ml, using specimens from HBV-positive patients.ResultsThe calculated LoD via a probit curve was 2.7 IU/ml for cobas 6800/8800 HBV, 2.8 IU/ml for cobas 4800 HBV, 9.6 IU/ml for CAP/CTM HBVv2, and 6.2 IU/ml for HPS/CTM HBVv2. The average accuracy was comparable between cobas 6800/8800 HBV, cobas 4800 HBV and CAP/CTM HBVv2 (0.04–0.05 log10 IU/ml), while a slightly lower accuracy was documented for HPS/CTM HBVv2 (−0.16 log10 IU/ml). A total of 211–245 clinical samples were used for a pairwise comparison. Mean paired log differences ranged from −0.17 log10 IU/ml to −0.01 log10 IU/ml. Coefficient of determination was over 98% for all pairs with high overall percent agreement at the 2000 and 20,000 IU/ml cutoffs (from 91.7% to 96.3%). In a subset of samples with VL±0.5 log10 to the 2000 and 20,000 IU/ml thresholds, concordance was still 72% and 82%, respectively.ConclusionsThe new cobas 6800/8800 HBV and 4800 HBV assays show high accuracy in samples with low-level viremia and a high concordance with the established HBV tests, CAP/CTM HBVv2 and HPS/CTM HBVv2, at 2000 and 20,000 IU/ml. Thus, all four HBV assays have high commutability and may be used interchangeably in routine clinical practice.

The Clinical Utility of Plasma Epstein–Barr Virus DNA Assays in Nasopharyngeal Carcinoma

The Clinical Utility of Plasma Epstein–Barr Virus DNA Assays in Nasopharyngeal Carcinoma

The clinical utility of plasma Epstein-Barr virus DNA assays in nasopharyngeal carcinoma: the dawn of a new era?: a systematic review and meta-analysis of 7836 cases.

In this study, we assessed the potential of plasma Epstein–Barr virus (EBV) DNA assays to predict clinical outcomes in a large sample of nasopharyngeal carcinoma (NPC) patients and proposed a risk stratification model based on standardized EBV DNA load monitoring.We conducted a meta-analysis of 14 prospective and retrospective comparative studies (n = 7 836 patients) to evaluate the correlation between pretreatment plasma EBV DNA (pre-DNA), midtreatment plasma EBV DNA (mid-DNA), posttreatment plasma EBV DNA (post-DNA), the half-life value of plasma EBV DNA clearance rate (t1/2), and clinical outcomes. Our primary endpoint was overall survival (OS). Our secondary endpoints were progression-free survival (PFS), distant-metastasis-free survival (DMFS), and local-regional-failure-free survival (LRFS).High pre-DNA, detectable mid-DNA, detectable post-DNA, and slow EBV DNA clearance rates were all significantly associated with poorer OS, with hazard radios (HRs) equal to 2.81, 3.29, 4.26, and 3.58, respectively. Pre-DNA, mid-DNA, and post-DNA had the same effects on PFS, DMFS, and LRFS.Plasma EBV DNA assays are highly prognostic of long-term survival and distant metastasis in NPC patients. Based on the results of this meta-analysis, we propose a 4-grade systematic risk stratification model. Given the inherent limitations of the included studies, future well-designed randomized clinical trials are required to confirm to the findings of this analysis and to contribute to the development of individualized treatment strategies for NPC patients.

Systemic treatment strategies and therapeutic monitoring for advanced nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is endemic in Asia and is etiologically associated with the Epstein–Barr virus. Although radiotherapy can cure most patients with early-stage disease, those with advanced disease often develop recurrences following radiotherapy. The past decade has witnessed significant advances in the systemic treatment of advanced NPC. This article reviews the latest literature regarding: combined modality therapy for locoregionally advanced NPC; clinical trials of novel biologic and cytotoxic therapies for metastatic NPC; and disease monitoring with the plasma Epstein–Barr virus DNA assay.

Failure of neonatal hepatitis B vaccination: the role of HBV-DNA levels in hepatitis B carrier mothers and HLA antigens in neonates

In a hepatitis B vaccination program (1982-1992), 705 infants born to HBsAg-positive mothers received HBIg within 2 h of birth and were vaccinated according to a three- or four-dose vaccination schedule, starting either at 3 months or directly after birth. Eight children HBsAg-positive during the first year of life (group 1: infected nonresponders). To determine whether failure of the hepatitis B vaccination was due to perinatal high-level maternal viraemia or genetically determined infant nonresponsiveness to the vaccine, we measured HBsAg and anti-HBs levels in infants and HBeAg and hepatitis B virus-DNA levels in maternal serum, and determined the HLA type of the infants. Controls included 14 infants with a normal anti-HBs response 1 year after vaccination (group 2: noninfected responders) and all eight infants without HBsAg and anti-HBs 1 year after vaccination (group 3: noninfected low responders). HBsAg, HBeAg and anti-HBs were measured by radioimmunoassay (Abbott Laboratories), hepatitis B virus-DNA was measured quantitatively by solution hybridization for groups 1, 2, and 3 (Abbott hepatitis B virus-DNA assay, Abbott Laboratories), and HLA was characterized by microcytotoxicity test for groups 1 and 3. All infants in groups 1 and 2 were born to HBeAg carrier mothers, and those in group 3 to HBeAg-negative mothers. Hepatitis B virus-DNA levels in maternal serum in group 1 were significantly higher than in group 2 (Wilcoxon rank-sum test: p < 0.01). Hepatitis B virus-DNA was not observed in group 3 maternal serum samples.(ABSTRACT TRUNCATED AT 250 WORDS)