scholarly journals ELISA versus Nested PCR for Detection of Hepatitis B Co-infection in HIV-infected People in Bandung Indonesia

2021 ◽  
pp. 991-1001
Author(s):  
Patricia Gita Naully
Keyword(s):  
Hepatitis B ◽  
Nested Pcr ◽  
False Positive ◽  
Hepatitis B Surface Antigen ◽  
False Negative ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hiv Patients ◽  
Nested Polymerase Chain Reaction ◽  
Pcr Method ◽  
Sensitivity Specificity

The number of Human Immunodeficiency Virus infected patients in Bandung is increasing. HIV patients are often suffering from Hepatitis B Virusco-infection. The co-infection status must be detected using the accurate method. This study aims to compare the accuracy of Enzyme-Linked Immunosorbent Assay and nested Polymerase Chain Reaction in detecting HIV-HBV co-infection in Bandung. The research method used is experimental. The sample used in this study was 50 people. All samples had met the inclusion criteria. The presence of hepatitis B surface antigen in the serum was detected using a qualitative ELISA Wantai kit, while sHBsAg gene in the whole blood was amplified using 2 pairs of the specific primary. The results obtained from both methods were analyzed to determine sensitivity, specificity, and accuracy values. The result of Hepatitis B detection using ELISA method showed that 15 samples (30%) were positive, 1 sample (2%) was a false positive, and 3 samples (6%) were the false negative. Moreover, the nested PCR method showed that 18 samples (36%) were positive without false positive or false negative result. These results showed that ELISA has a sensitivity value of 83.33%, specificity of 96.87%, and the accuracy of 92%, while Nested PCR has a value of 100% for all three parameters. Therefore, it can be concluded that the Nested PCR method was more accurate than ELISA to detect Hepatitis B co-infection in HIV patients in Bandung.The Nested PCR can be recommended for detecting the other co-infection cases.   Keywords: accuracy, HBsAg, co-infection, sensitivity, specificity

scholarly journals Study on the Prevalence of Occult Hepatitis B Virus Infection in Patients Undergoing Hemodialysis

2020 ◽  
Author(s):  
Shahab Mahmoudvand ◽  
Somayeh Shokri ◽  
Habibollah Mirzaei ◽  
Manoochehr Makvandi ◽  
Ali Teimoori ◽  
...  
Keyword(s):  
Hepatitis B ◽  
Nested Pcr ◽  
Hepatitis B Surface Antigen ◽  
Hbv Dna ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peripheral Blood Mononuclear ◽  
Cross Sectional ◽  
Occult Hepatitis ◽  
Occult Hepatitis B ◽  
Sera Samples

Occult hepatitis B (OBI) is a major challenging clinical entity characterized by the absence of hepatitis B surface antigen (HBsAg). The persistence of OBI may progress to fibrosis, cirrhosis, and hepatocellular carcinoma. This study was aimed to investigate the prevalence of OBI among HD patients. In the present cross-sectional study, 89 sera samples of hemodialysis individuals were tested for HBsAg and HBcIgG by Enzyme-linked Immunosorbent Assay (ELISA). In addition, the HBV DNA has tested in sera and peripheral blood mononuclear cell (PBMC) samples by nested-PCR. Out of 89 patients, 51(57.3%) were males, and 38 (42.7%) females. The ages ranged from 24 to 90 years (with a mean of 57.5±1.37 years). All the sera samples had normal levels of Aspartate Aminotransferase (AST) and Alanine Transaminase (ALT) but had high levels of Creatinine (Cr) (6.9±2.17) and Blood Urea Nitrogen (BUN) (61.83±2.03). 2/89 (2.2%) sera samples were positive for both HBsAg and HBc-IgG test; in addition, HBV DNA was detected in both sera and their PBMC samples. The sera of 15/89 (16.85%) were only positive for the HBc-IgG test, including 10/51 (19.6%) males and 5/38 (13.2%) females (P=0.5). The high 16.85% prevalence OBI has been found among HD patients. To manage OBI infection, screening of HBV DNA should be implemented for HD patients by sensitive molecular means such as nested-PCR and real-time PCR.

Need for Confirmatory Neutralization Tests for Hepatitis B Surface Antigen Tests in Populations with Intermediate Prevalence

2021 ◽  
Author(s):  
Min Young Lee ◽  
So Young Kang ◽  
Woo In Lee ◽  
Myeong Hee Kim
Keyword(s):  
Hepatitis B ◽  
Surface Antigen ◽  
Neutralization Test ◽  
Hepatitis B Surface Antigen ◽  
False Negative ◽  
Confirmatory Test ◽  
Negative Group ◽  
Positive Group ◽  
Test Kit ◽  
Antigen Tests

Abstract Objective Hepatitis B surface antigen (HBsAg) is known as the hallmark of hepatitis B virus (HBV) infection. This study aimed to determine whether an HBsAg neutralization test is necessary to accurately interpret HBsAg test results. Methods Initially reactive HBsAg specimens from a 5-year period, with cutoff index values between 1.0 and 2.0, were subjected to neutralization confirmatory testing using an Elecsys HBsAg Confirmatory test kit (Roche Diagnostics GmbH. Mannheim, Germany). Results The neutralization test showed 46.1% positive (confirmed positive group) and 53.9% negative (confirmed negative group) results from the total specimens. Among the confirmed negative group, 79.5% of patients were confirmed to be negative for the current infection, whereas 4 patients in the chronic hepatitis B subgroup showed a neutralization percentage close to 40%. More than half of patients in the confirmed positive group were considered to be in the hepatitis B e antigen-negative inactive HBsAg carrier phase. Conclusion In populations with intermediate HBV prevalence, a neutralization test is necessary to confirm an HBsAg result and reduce the false positive and false negative rates of initial HBsAg tests.

scholarly journals Evaluation of optimal urine screening and confirmation cut-off values for opiates, at a national reference laboratory

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Çiğdem Karakükcü ◽  
Mehmet Zahid Çıracı ◽  
Derya Kocer ◽  
Mine Yüce Faydalı ◽  
Muhittin Abdulkadir Serdar
Keyword(s):  
False Positive ◽  
False Positive Rate ◽  
False Negative ◽  
Urine Samples ◽  
Reference Laboratory ◽  
National Reference Laboratory ◽  
Urine Screening ◽  
National Reference ◽  
Positive Rate ◽  
Sensitivity Specificity

Abstract Objectives To obtain optimal immunoassay screening and LC-MS/MS confirmation cut-offs for opiate group tests to reduce false positive (FP) and false negative (FN) rates. Methods A total of 126 urine samples, −50 opiate screening negative, 76 positive according to the threshold of 300 ng/mL by CEDIA method – were confirmed by a full-validated in-house LC-MS/MS method. Sensitivity, specificity, FP, and FN rates were determined at cut-off concentrations of both 300 and 2,000 ng/mL for morphine and codeine, and 10 ng/mL for heroin metabolite 6-mono-acetyl-morphine (6-MAM). Results All CEDIA opiate negative urine samples were negative for morphine, codeine and 6-MAM. Although sensitivity was 100% for each cut-off; specificity was 54.9% at CEDIA cut-off 300 ng/mL vs. LC-MS/MS cut-off 300 ng/mL and, 75% at CEDIA cut-off 2,000 ng/mL vs. LC-MS/MS cut-off 2,000 ng/mL. False positive rate was highest (45.1%) at CEDIA cut-off 300 ng/mL. At CEDIA cut-off 2,000 ng/mL vs. LC-MS/MS cut-off 300 ng/mL, specificity increased to 82.4% and FP rate decreased to 17.6%. All 6-MAM positive samples had CEDIA concentration ≥2,000 ng/mL. Conclusions 2,000 ng/mL for screening and 300 ng/mL for confirmation cut-offs are the most efficient thresholds for the lowest rate of FP opiate results.

scholarly journals Significant Increase of HBV Infection in HIV Infected Patients With Disease Progression in China Based on Two MSM HIV Infected Cohorts

2021 ◽  
Author(s):  
Zhiqiang Zhu ◽  
Qi Liang ◽  
Taiyi Jiang ◽  
Yanmei Jiao ◽  
Yu Zhang
Keyword(s):  
Hepatitis B ◽  
Hiv Infection ◽  
Disease Progression ◽  
Hepatitis B Surface Antigen ◽  
Chronic Phase ◽  
Hbv Infection ◽  
Hiv Patients ◽  
Acute Hiv Infection ◽  
Specific Antibodies ◽  
Acute Hiv

Abstract The date about the condition of HBV co infection with the disease progress of HIV is limited. To investigate whether the incidence of HBV co-infection is significantly higher in HIV patients with disease progression in China, we compared rates of HBV co-infection in HIV patients based on an acute and a chronic HIV infected cohort. Significance was assessed with Chi-square. HBV infection is diagnosed by the presence of hepatitis B surface antigen. The HBsAg positive rate increased from 6.18% in acute HIV infection to 11.44% in chronic HIV infection. Thirty-four acute HIV patients had been tested for HBV in their chronic phase, four of them had HBV -specific antigens and/or specific antibodies changes. The number of Hepatitis B virus-specific antibodies decreased from acute phase to chronic phase in four patients and two patients’ HBsAg changed from negative to positive. There is an increased prevalence of HBV infection in HIV patients with the disease progression in China.

scholarly journals Occult hepatitis B virus infection among hemodialysis patients

2018 ◽  
Vol 91 (2) ◽  
Author(s):  
Rahil Nahid Samiei ◽  
Somayeh Shokri ◽  
Shahab Mahmoudvand ◽  
Manoochehr Makvandi ◽  
Heshmatollah Shahbazian ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hbv Dna ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hepatitis B Infection ◽  
Nested Polymerase Chain Reaction ◽  
Occult Hepatitis ◽  
B Virus ◽  
The World ◽  
Occult Hepatitis B

Hepatitis B virus is a major public health impasse all over the world. Recently a new form of hepatitis B infection named Occult hepatitis B Infection (OBI) has appeared globally. The OBI is defined as the presence of HBV DNA in the liver and/or blood in the absence of detectable serum HBsAg with/without anti-HBc or anti-HBs. The prevalence of OBI has been reported in hemodialysis (HD) patients in different regions of the world. Thus, this study investigated the prevalence of OBI among HD patients. The cross-sectional study was carried out on 84 HD patients. These sera were checked for HBsAg, HBc-IgG assessment using Enzyme linked immunosorbent assay. The DNA was extracted from the sera samples and tested for HBVDNA detection using Nested Polymerase Chain Reaction (Nested PCR). The liver function tests including serum alanine aminotransferase and aspartate aminotransferase levels were carried out for all the HD individuals. 52/84(61.9%) of HD were males and 32/84 (38.1%) were females. The patient’s age ranged from 25 to 64 with a mean age of 52.4±15.2 years. HBsAg and HBc-IgG were detected in 1(1.1%) female. 2 (2.4%; a female and a male) patients were positive for HBsAg. 14/84 (16.7%; 6 female and 8 male) HD patients were positive for anti-HBc but negative for HBsAg, among them 4(28.6%; 2 female and 2 male) cases were positive for HBV DNA, indicating the presence of OBI in HD patients. Even distribution of OBI among the HD was found in 2(2.36%) male and 2(2.36%) female (P>.0.05). In the present study the moderate rate of 4.76% OBI has been observed in HD patients. The prevalence of seropositive OBI among the gender was 2(2.36%) male and 2(2.36%) female. The seronegative OBI have not been detected in the present study but requires further investigation. In this study the affliction of OBI in HD patients is not clear.

scholarly journals Expression Level of Small Envelope Protein in Addition to Sequence Divergence inside Its Major Hydrophilic Region Contributes to More Efficient Surface Antigen Secretion by Hepatitis B Virus Subgenotype D2 than Subgenotype A2

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 967
Author(s):  
Qianru Wang ◽  
Shuwen Fu ◽  
Jing Zhang ◽  
Quan Yuan ◽  
Jisu Li ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Envelope Protein ◽  
Hepatitis B Surface Antigen ◽  
Site Directed Mutagenesis ◽  
Enzyme Linked Immunosorbent Assay ◽  
S Protein ◽  
Huh7 Cells ◽  
B Virus

Hepatitis B surface antigen (HBsAg) promotes persistent hepatitis B virus (HBV) infection. It primarily corresponds to small (S) envelope protein secreted as subviral particles. We previously found that genotype D clones expressed less S protein than genotype A clones but showed higher extracellular/intracellular ratio of HBsAg suggesting more efficient secretion. The current study aimed to characterize the underlying mechanism(s) by comparing a subgenotype A2 clone (geno5.4) with a subgenotype D2 clone (geno1.2). Five types of full-length or subgenomic constructs were transfected to Huh7 cells at different dosage. HBsAg was quantified by enzyme linked immunosorbent assay while envelope proteins were detected by Western blot. We found that ratio of extracellular/intracellular HBsAg decreased at increasing amounts of DNA transfected. Conflicting findings from two types of subgenomic construct confirmed stronger secretion inhibitory effect of the genotype D-derived large envelope protein. Chimeric constructs followed by site-directed mutagenesis revealed geno1.2 specific V118/T127 and F161/A168 in the S protein as promoting and inhibitory of HBsAg secretion, respectively. In conclusion, more efficient HBsAg secretion by subgenotype D2 than subgenotype A2 is attributed to lower level of S protein expression in addition to V118 and T127 in S protein, although its F161 and A168 sequences rather reduce HBsAg secretion.

scholarly journals Development and Technical and Clinical Validation of a Quantitative Enzyme-Linked Immunosorbent Assay for the Detection of Human Antibodies to Hepatitis B Surface Antigen in Recipients of Recombinant Hepatitis B Virus Vaccine

2009 ◽  
Vol 16 (8) ◽  
pp. 1236-1246 ◽  
Author(s):  
Pierre Cambron ◽  
Jeanne-Marie Jacquet ◽  
Bernard Hoet ◽  
Marc Lievens
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Immunosorbent Assay ◽  
Surface Antigen ◽  
Hepatitis B Surface Antigen ◽  
Geometric Mean ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Limit Of Quantitation ◽  
B Virus

ABSTRACT Pending removal from the market of a commercial assay (the AUSAB [Abbott Laboratories] enzyme immunoassay [EIA]) for the determination of antibodies to hepatitis B surface antigen (HBsAg), a new in-house quantitative enzyme-linked immunosorbent assay (ELISA) to measure antibodies against HBsAg (anti-HBs) was developed (anti-HBs in-house). Specific anti-HBs antibodies were sandwiched between the precoated HBsAg ad and ay subtypes purified from plasma from hepatitis B virus (HBV) human carriers and the recombinant HBsAg adw2 subtype tagged with horseradish peroxidase. The assay was calibrated against the 1st International Reference Preparation for anti-hepatitis B immunoglobulin (lot 1977-W1042). Analytical sensitivity and the limit of quantitation were estimated at 0.43 mIU/ml and 2.0 mIU/ml, respectively. Overall reproducibility was 11.86%, and accuracy was estimated to be 94.89%. More than 4,000 samples from seven clinical trials were tested with the anti-HBs in-house assay and compared to results generated with AUSAB EIA and AUSAB radioimmunoassay (RIA). During the technical validation, the anti-HBs in-house assay was compared to the AUSAB RIA as a reference (n = 919). Overall assessment of concordance and Deming's regression analysis were performed. The coefficient of correlation between the AUSAB RIA and anti-HBs in-house assay was 0.9815 with a slope of 0.9187. The overall agreement between anti-HBs in-house and AUSAB RIA was 97.61%, considering the clinical cutoffs at 3.3 mIU/ml and 1.0 mIU/ml for the respective assays. From a clinical perspective, seroprotection rates and anti-HBs geometric mean antibody concentrations for individual studies calculated with either the in-house assay or the reference assays were similar. Conclusions of individual studies were confirmed. The performance characteristics of the in-house assay are acceptable. There is no evidence that use of the new assay would lead to different clinical conclusions from the reference method.

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