Hepatitis B Virus Detection Research Articles

One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications.

Viral hepatitis caused by hepatitis B virus (HBV) and hepatitis C virus (HCV) infections remain a major global public health challenge, particularly in low- and middle-income countries. It is crucial to utilize a pointof-care (POC) testing platform that is sensitive, specific, rapid, and user-friendly for screening and diagnosis of the two infections. Here, a novel molecular diagnostic assay, integrating multiplex loop-mediated isothermal amplification with a gold nanoparticle-based lateral flow biosensor (mLAMP-AuNPs-LFB) was developed and applied for one-step, visual, rapid, sensitive, and specific identification of HBV and HCV. The AuNPs-based LFB was devised and constructed for the simultaneous detection of HBV and HCV. The HBV-LAMP and HCV-LAMP primers were designed against the S and 5'-untranslated region (5'-UTR) genes from the major HBV genotypes (B, C, D, B/C recombinant, and C/D recombinant) and HCV subtypes (1b, 2a, 3a, 3b, and 6a) in China, respectively. Our assay conditions, both multiplex-LAMP amplification temperature and time were optimized. The sensitivity and specificity of our assay were tested, and the feasibility of our assay was verified through clinical samples. The AuNPs-based LFB used here was successfully manufactured according to our devise manual. The two unique independent primer pairs were successfully designed based on the S and 5'-UTR genes, respectively. The optimal mLAMP-AuNPs-LFB detection process, involving rapid nucleic acid isolation (10min), mLAMP (63°C for 35min), and visual AuNPs-LFB interpretation (less than 2min), could be completed within 50min. The HBV&HCV-mLAMP-AuNPs-LFB assay can detect the target genes (HBV-S and HCV-5'-UTR) with as low as 20 copies of plasmid template per test, and the specificity was 100% for the experimental pathogens. The preliminary results manifested that our mLAMP-AuNPs-LFB assay is a valuable tool and has tremendous potential as a POC testing approach for HBV and HCV identification, especially in undeveloped regions.

Incidence of Occult Hepatitis B Infection (OBI) and hepatitis B genotype characterization among blood donors in Cameroon.

Occult hepatitis B infection (OBI) is characterized by the presence of hepatitis B virus (HBV) DNA at low levels in serum (<200 IU/mL) with a negative hepatitis B surface antigen (HBsAg) test. OBI remains a major challenge to blood safety, particularly in HBV-endemic regions like Cameroon, where HBV detection relies solely on HBsAg testing. This cross-sectional study aimed to investigate the actual incidence and genotype characteristics of OBI in Cameroonian blood donors. Between March and June 2023, samples were collected from 288 HBsAg-negative blood donors aged 18 to 55 years and analysed for antibodies against the HBV core (anti-HBc) and surface antigens (anti-HBs). Following DNA extraction from the serum samples, qualitative nested PCR and quantitative real-time PCR were used to detect HBV viral DNA and viral load respectively. For positive samples, sequencing of a fragment of the S gene was performed to identify the circulating HBV genotypes. The findings revealed that 58% (n = 167/288) of blood donors tested positive for anti-HBc, 29% (n = 83/288) tested positive for anti-HBs, and 26% (n = 75/288) being positive for both anti-HBc and anti-HBs. Occult hepatitis was confirmed in 4.5% of the blood donors, all of whom belonged to either HBV genotypes A or E, which are predominant in Cameroon. The amino acid substitution sA184V associated with HBsAg detection failure in genotype E was observed in 70% of OBI sequences, and the HBsAg immune escape variants (sT131N and sS143L) implicated in OBI were also observed. The mutation rtN139K in the reverse transcriptase (RT) domain of the overlapping HBV polymerase (P) gene was present in 17% of OBI-positive sequences of genotype E, likely contributing to masking HBsAg secretion. The results suggest a considerable risk of transfusion-transmitted HBV in this region. Therefore, to ensure blood safety, nucleic acid testing (NAT) is recommended, as relying solely on HBsAg assays is insufficient to eliminate this risk.

Impact of universal hepatitis B virus (HBV) screening using chemotherapy orders on the HBV reactivation in cancer patients.

Hepatitis B virus (HBV) reactivation (HBVr) induced by chemotherapy in patients with resolved or chronic infection can lead to severe consequences. Despite recommendations, rates of HBV screening before chemotherapy are low due to poor recognition of risk factors by clinicians. The aim of the study is to assess whether routine HBV screening using universal HBV screening on chemotherapy orders (CO) could reduce HBVr incidence. This is a 1-year retrospective single-center observational study of patients who received intravenous chemotherapy post implementation of CO. We compared the incidence of HBVr in three groups of patients: those screened through CO (group 1), those screened by the medical team (group 2), and those not screened (group 3). On a total of 1374 patients, 179 of 206 patients were screened as requested on CO (group 1) and 421 by the medical team (group 2), whereas 747 patients were not screened (group 3). Only one HBVr occurred, and no difference was seen on the incidence of HBVr between group 1 and group 3 (0% vs 0.1%; p = 1.00), probably because of a lack of follow-up after chemotherapy. Follow-up for HBVr was imperfect in group 1 and group 2 (16.7% vs 5.6%; p = 0.32). Screening was done for 92% of patients on anti-CD20 therapy. In group 3, 89 patients had ALT elevation during chemotherapy but only 17 (19%) were tested for HBVr. Systematic HBV detection requested on CO is an effective way to obtain a high percentage of patients with adequate screening, particularly when chemotherapy is at high risk of HBVr. Nevertheless, this screening method do not guarantee optimal follow-up and requires improvements.

Seroprevalence of Hepatitis B Virus among Blood Donors at National Blood Transfusion and Research Center-Taiz Branch, Yemen

Hepatitis B Virus (HBV) represents one of the most health problems. Almost more than 350 million infected persons were reported universally. Blood transfusion is commonly the most common route of transmission of HBV. Recently, Hospitals in Yemen are increasing demands for blood and blood products due to war injury, anemia, and malnutrition. The risk of transfusion transmission of infection relatively increases. This study aimed to determine hepatitis B virus prevalence among blood donors at the National Blood Transfusion Center -Taiz branch (NBTRC- TB). Data was collected from the National Blood Transfusion and Research Center (Taiz branch) from 1/4/2020 to 16/12/2020. A total of 3174 blood donors donated blood at the center. A descriptive cross-sectional study was conducted on them. Blood donor information was registered at the reception department, and all blood screening tests were performed. Data of HBsAg and total anti-HBc were analyzed statistically by using IBM SPSS version 26. Among 3174 blood donors, male blood donors were 3158 (99.5%) and female blood donors 16 (0.5%). Furthermore, 41 (1.3%) were positive for HBsAg (which all the positive results were for male donors and no positivity were for female donors) while 353 (11.1%) were positive for total anti- HBc Ab which is distributed at ratio of 11% (350) for males and 0.1% (3) for females. On the other hand, the result of HBsAg and total anti-HBc Ab together showed three levels of positivity as the (HBsAg positive / anti-HBc negative) were 12 (0.4%) in which was a low percentage and reversely, the (HBsAg negative/ HBcAb positive) were 316 (10.2%), While the HBsAg positive / anti-HBc positive were 27 (0.9%). So this study shed light on the result of HBsAg negative/ anti-HBc positive which the percentage (10.2%) of which statistically significant (p. value = 0.000) and was the highest result indicates donors with post HBV infection or have low HBV infection. This study showed the high prevalence of post-HBV infection among blood donors. Using both markers HBsAg and total anti-HBc Ab will improve the detection of HBV before blood transfusion.

Systematic Optimization of Universal Real-Time Hypersensitive Fast Detection Method for HBsAg in Serum Based on SERS.

Hepatitis B virus (HBV) is a major cause of liver cirrhosis and hepatocellular carcinoma, with HBV surface antigen (HBsAg) being a crucial marker in the clinical detection of HBV. Due to the significant harm and ease of transmission associated with HBV, HBsAg testing has become an essential part of preoperative assessments, particularly for emergency surgeries where healthcare professionals face exposure risks. Therefore, a timely and accurate detection method for HBsAg is urgently needed. In this study, a surface-enhanced Raman scattering (SERS) sensor with a sandwich structure was developed for HBsAg detection. Leveraging the ultrasensitive and rapid detection capabilities of SERS, this sensor enables quick detection results, significantly reducing waiting times. By systematically optimizing critical factors in the detection process, such as the composition and concentration of the incubation solution as well as the modification conditions and amount of probe particles, the sensitivity of the SERS immune assay system was improved. Ultimately, the sensor achieved a sensitivity of 0.00576 IU/mL within 12 min, surpassing the clinical requirement of 0.05 IU/mL by an order of magnitude. In clinical serum assay validation, the issue of false positives was effectively addressed by adding a blocker. The final sensor demonstrated 100% specificity and sensitivity at the threshold of 0.05 IU/mL. Therefore, this study not only designed an ultrasensitive SERS sensor for detecting HBsAg in actual clinical serum samples but also provided theoretical support for similar systems, filling the knowledge gap in existing literature.

Development of Molecular-Based Screening Test for Hepatitis B Virus in Human Plasma Samples.

Despite regular administration of hepatitis B virus (HBV) vaccine in several countries, the mortality rate associated with HBV remains significant. The antiviral medications available for the treatment of HBV infection do not suffice for the serious complications related to chronic HBV infection. Additionally, the serological tests fail to detect early viral replication preventing early treatment response. Recently, many studies have demonstrated the significant advantages of loop-mediated isothermal amplification (LAMP) over serological testing and polymerase chain reaction (PCR), for the rapid detection of microbial pathogens. Here we developed a rapid, sensitive, and portable system-integrative LAMP assay for the detection of HBV DNA in plasma samples. The final optimized assay was achieved with an amplification time of less than 45 min at 62°C. The assay showed 100% specificity, 92.20% sensitivity, and a detection limit of 10 copies/µL in 77 HBV-positive plasma samples with known Cq values. Our results showed that the colorimetric LAMP assay is sensitive, efficient, and supremely reliable for rapid detection of HBV, and may be potentially used as a screening test in areas with poor laboratory facilities and limited resource availability.

Development of a multiplex polymerase chain reaction assay for detection of hepatitis C virus, hepatitis B virus, and human immunodeficiency virus 1.

Hepatitis C virus (HCV), hepatitis B virus (HBV), and human immunodeficiency virus 1 (HIV-1) are the most epidemic blood-borne viruses, posing threats to human health and causing economic losses to nations for combating the infection transmission. The diagnostic methodologies that depend on the detection of viral nucleic acids are much more expensive, but they are more accurate than serological testing. To develop a rapid, cost-effective, and accurate diagnostic multiplex polymerase chain reaction (PCR) assay for simultaneous detection of HCV, HBV, and HIV-1. The design of the proposed PCR assay targets the amplification of a short conserved region featured with a distinguishable melting profile and electrophoretic molecular weight inside each viral genome. Therefore, this diagnostic method will be appropriate for application in both conventional (combined with electrophoresis) and real-time PCR facilities. Confirmatory in silico investigations were conducted to prove the capability of the approached PCR assay to detect variants of each virus. Then, Egyptian isolates of each virus were subjected to the wet lab examination using the given diagnostic assay. The in silico investigations confirmed that the PCR primers can match many viral variants in a multiplex PCR assay. The wet lab experiment proved the efficiency of the assay in distinguishing each viral type through high-resolution melting analysis. Compared to related published assays, the proposed assay in the current study is more sensitive and competitive with many expensive PCR assays. This study provides a simple, cost-effective, and sensitive diagnostic PCR assay facilitating the detection of the most epidemic blood-borne viruses; this makes the proposed assay promising to be substitutive for the mistakable and cheap serological-based assays.

Dual Detection of Hepatitis B and C Viruses Using CRISPR‐Cas Systems and Lateral Flow Assay

The development of sensitive and specific diagnostic tools for hepatitis B virus (HBV) and hepatitis C virus (HCV) remains crucial for effective disease management and control. In this study, we utilized CRISPR‐Cas12 and CRISPR‐Cas13 systems for the detection of HBV (DNA virus) and HCV (RNA virus), respectively. We designed and tested multiple guide RNAs (gRNAs) targeting both viruses, confirming successful cleavage of target sequences through gel electrophoresis and a fluorescent reporter assay. Using optimized gRNAs, we developed a lateral flow assay (LFA) for sensitive detection of HBV and HCV, demonstrating a concentration‐dependent signal increase. Importantly, no cross‐reactivity was observed with other viral targets. To further enhance sensitivity, we employed a dual‐enzyme approach, combining Cas12 and Cas13 in a single reaction, which significantly improved detection limits for both viruses. Finally, we developed a dual antigen detection LFA strip capable of simultaneously detecting both HBV and HCV in a single sample. This approach holds promise for point‐of‐care (POC) diagnostics where the specific viral infection is unknown. This study addresses the current limitations in CRISPR‐Cas based diagnostics, namely, the need for ultrasensitive detection methods and the ability to detect multiple antigens using a single test strip. Our findings demonstrate the feasibility of using CRISPR‐Cas systems for highly sensitive and specific detection of HBV and HCV, paving the way for potential POC application.

Metatranscriptomic data mining together with microfluidic card uncovered the potential pathogens and seasonal RNA viral ecology in a drinking water source.

Despite metatranscriptomics becoming an emerging tool for pathogen surveillance, very little is known about the feasibility of this approach for understanding the fate of human-derived pathogens in drinking water sources. We conducted multiplexed microfluidic cards and metatranscriptomic sequencing of the drinking water source in a border city of North Korea in four seasons. Microfluidic card detected norovirus, hepatitis B virus (HBV), enterovirus, and Vibrio cholerae in the water. Phylogenetic analyses showed that environmental-derived sequences from norovirus GII.17, genotype C of HBV, and coxsackievirus A6 (CA6) were genetically related to the local clinical isolates. Meanwhile, metatranscriptomic assembly suggested that several bacterial pathogens, including Acinetobacter johnsonii and V. cholerae might be prevalent in the studied region. Metatranscriptomic analysis recovered 349 species-level groups with substantial viral diversity without detection of norovirus, HBV, and CA6. Seasonally distinct virus communities were also found. Specifically, 126, 73, 126, and 457 types of viruses were identified in spring, summer, autumn, and winter, respectively. The viromes were dominated by the Pisuviricota phylum, including members from Marnaviridae, Dicistroviridae, Luteoviridae, Potyviridae, Picornaviridae, Astroviridae, and Picobirnaviridae families. Further phylogenetic analyses of RNA (Ribonucleic Acid)-dependent RNA polymerase (RdRp) sequences showed a diverse set of picorna-like viruses associated with shellfish, of which several novel picorna-like viruses were also identified. Additionally, potential animal pathogens, including infectious bronchitis virus, Bat dicibavirus, Bat nodavirus, Bat picornavirus 2, infectious bursal disease virus, and Macrobrachium rosenbergii nodavirus were also identified. Our data illustrate the divergence between microfluidic cards and metatranscriptomics, highlighting that the combination of both methods facilitates the source tracking of human viruses in challenging settings without sufficient clinical surveillance.

Development of a Reagent Kit for the Quantitative Determination of Hepatitis B Virus (HBV) DNA in Clinical Material by PCR with Hybridization-Fluorescence Detection

Relevance.Hepatitis B virus (HBV) is one of the most common viral infections affecting people worldwide and can lead to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Currently 3% of the world's population are infected with hepatitis B virus and are at risk of developing life-threatening liver disease. Immunological and molecular biological methods of detection of HBV are currently used in laboratory diagnostics. The polymerase chain reaction (PCR) is currently the most sensitive method for the detection and quantification of HBV. HBV DNA quantification is widely used to monitor the antiviral treatment of HBV infection.Aim.To develop a real-time PCR kit for the quantification of HBV DNA.Materials and methods.A total of 200 plasma and serum samples positive and negative for HBV were used in the development. The performance of the developed kit was compared with the use of other commercially registered HBV diagnostic kits in Russia. Additionally, the nucleotide sequences of all existing virus genotypes analysed for the selection of primers using GeneBank system.Results and discussion.Comparison analysis of the results of quantitative determination by real-time PCR in 200 clinical serum and blood plasma samples showed that the diagnostic sensitivity of the developed kit was 100% and specificity 100%. The primers developed specific to the POL gene region. The kit is capable of detecting all types of virus genotypes.Conclusions.The developed reagent kit allows detection of hepatitis B virus and determination of its quantity within 70 minutes. In addition to a large number of genotypes and subgenotypes, the virus is characterized by mutational changes in the genome, which complicates its diagnosis and, as a consequence, the ongoing therapy with drugs. Conservative regions for primer and probe selection taken into account in the development, and the sequencing results obtained are applicable to all HBV genotypes. The reagent kit is designed to monitor HBV infected patients and will allow the analysis of different HBV viral loads.

A multifunctional photoelectrochemical biosensing platform based on CdS QDs@TiO2-sensitizing Ni-TBAPy-SC coupled with multiple amplification for detection of HBV and thrombin

A multifunctional photoelectrochemical (PEC) biosensor using CdS quantum dots (QDs)-sensitized nickel-metal organic frameworks (MOFs) coupled with multiple amplification techniques was developed for detection of hepatitis B virus (HBV) and thrombin (TB). The tetra (p-benzoate) pyrene (TBAPy)Ni-SC was firstly modified on the electrode and showed an ultra-stable anodic photocurrent with excellent reproducibility. Subsequently AgInS2 QDs were attached to the electrode to much decrease the background signal and improved the assay sensitivity. Moreover, an effective PEC sensitizing probe was specifically designed by connecting a large number of CdS QDs onto TiO2 nanoparticles (NPs). After HBV target induced cyclic amplification to produce abundant output DNA, the CdS QDs@TiO2 signal probes were attached to the electrode by DNA hybridization, which turned the PEC signal of Ni-TBAPy-SC “on”, thus achieving sensitive detection of HBV. In addition, the CdS QDs@TiO2 nano-signal probe is released while target thrombin specifically binds to its aptamer, turning the PEC signal of Ni-TBAPy-SC “off” for thrombin detection. This work has developed a novel QD-sensitized Ni-TBAPy-SC PEC biosensor, which is able to rapidly detect dual targets and avoid false results, showing important value for early diagnosis of diseases.

Evaluation of three hepatitis b virus detection protocols in blood donors

Hepatitis B virus (HBV) DNA diagnosis plays a critical role in the transmission of HBV infections. Accurate protocols in HBV detection will reduce the spread of the virus from blood donors. Therefore, this research compared the HBV detecting potentials of three different HBV protocols in the virus detection. Blood from 200 donors at the University college blood bank Ibadan were sampled and tested for HBV using the ABON™ HBsAg Hepatitis B Surface Antigen Tests (strip test), Monolisa TM HBsAg ULTRA (ELISA test) and Huruian biology HBV detection kit (qPCR test). HBV positive samples were genotyped by sequencing to ascertain genotype identity. Results showed that of the 200 samples tested, both ABON™ HBsAg Hepatitis B Surface Antigen Tests and Monolisa TM HBsAg ULTRA (for ELISA) did not record any positive sample. However, Huruian biology HBV detection kit via qPCR recorded a total of twenty-three positive samples. Of the twenty-three samples, twenty were of genotype E while three were of genotype B. The results from this study showed that that qPCR is more accurate in HBV virus detection than ELISA technique and also HBV genotype E is the most common genotype isolated in Nigeria.

Prevalence of Hepatitis B, C and D Infections in Preoperative Patients at a Tertiary Hospital in Southern Nigeria

Patients going for surgical operations are required to be screened for hepatitis B virus (HBV) and hepatitis C virus (HCV) infections among other transmissible infections. This is necessary for better assessment of the patient's condition alongside the need for extra precautions during surgery. Hepatitis D virus (HDV) infection is not commonly reported but occurs in association with commonly reported but occurs in association with that of hepatitis B virus. The epidemiology of this co-infection is worth investigating, as it constitutes a more aggressive form of hepatitis Following due ethical considerations, this cross-sectional descriptive study enrolled 180 pre-operative patients for viral hepatitis screening. Immunochromatographic methods were used for detection of HBV and HCV infections, while detection of HDV was based on enzyme-linked immunosorbent assay method. Data processing was carried out using SPSS version 22.0. Results are presented as numbers and frequencies as percentage. The prevalence of HBV, HCV and HDV as recorded among pre-operative patients was 7.8%, 1.7% and 1.1% respectively. Hepatitis B virus alone accounted for 68.8% of all hepatitis cases, while its co-infection with HDV and HCV were observed to be 12.5% and 6.2% respectively Single infection of HCV was seen to be 12.5% of all infected cases. Hepatitis B virus was observed to be the most prevalent viral hepatitis agent within the studied population, while hepatitis D virus was detected at a rate of 1.1%

Development and clinical validation of a one-step pentaplex real-time reverse transcription PCR assay for detection of hepatitis virus B, C, E, Treponema pallidum, and a human housekeeping gene

BackgroundWith the safety of blood transfusion being a major public health concern, the development of a rapid, sensitive, specific, and cost-effective multiplex PCR assay for simultaneous detection of hepatitis B virus(HBV), hepatitis C virus (HCV), hepatitis E virus (HEV), and Treponema pallidum(T. pallidum) in blood is crucial.MethodsFive primer pairs and probes were designed towards conserved regions of target genes and used to establish a one-step pentaplex real-time reverse transcription PCR(qRT-PCR) assay for simultaneous detection of HBV, HCV, HEV, T. pallidum, and RNase P(housekeeping gene), providing sample quality check. The clinical performance of the assay was further determined with 2400 blood samples from blood donors and patients in Zhejiang province, and compared the results with commercial singleplex qPCR and serological assays.ResultsThe 95% limit of detection(LOD) of HBV, HCV, HEV, and T. pallidum were 7.11 copies/µL, 7.65 copies/µL, 8.45 copies/µL, and 9.06 copies/µL, respectively. Moreover, the assay has good specificity and precision. Compared to the singleplex qPCR assay, the novel assay for detecting HBV, HCV, HEV, and T. pallidum presented 100% clinical sensitivity, specificity, and consistency. Several discrepant results between serological and pentaplex qRT-PCR assays were found. Of 2400 blood samples, there were 2(0.08%) HBsAg positive samples, 3(0.13%) anti-HCV positive samples, 29(1.21%) IgM anti-HEV positive samples and 6(0.25%) anti-T. pallidum positive samples proven negative in nucleic acid detection. 1(0.04%) HBV DNA positive sample and 1(0.04%) HEV RNA positive sample were detected negative by serological testing.ConclusionsThe developed pentaplex qRT-PCR is the first assay on simultaneous, sensitive, specific, and reproducible detection of HBV, HCV, HEV, T. pallidum, and RNase P in a single tube. It could detect pathogens in blood during the window period of infection and is a good tool for effectively screening blood donors and early clinical diagnosis.

Highly Sensitive Hepatitis B Virus Identification by Antibody-Aptamer Sandwich Enzyme-Linked Immunosorbent Assay

Hepatitis B virus (HBV) infection is considered a major global health problem, causing various health issues, including cirrhosis, hepatitis, and liver cancer. The risk of developing such complications increases when the viral load is above 105 copies/mL. Early identification of HBV infection is imperative to preventing the spread of infection to other parts of the body. Although various sensing methods have been developed to identify HBV, researchers are still working toward developing cheap, easy, and sensitive detection methods. We developed a highly sensitive and rapid HBV detection method using nanomaterials on enzyme-linked immunosorbent assay (ELISA). Aptamer-antibody was utilized as the detection probe and immobilized on a zeolite-modified ELISA plate to detect the HBV biomarker hepatitis B surface antigen (HBsAg). To enhance the detection of HBsAg, aptamer and antibody were attached to gold nanoparticle through electrostatic interaction and immobilized on the zeolite-modified ELISA plate through amine linker. This probe-modified ELISA plate detected low levels of HBsAg, with a detection limit of 0.1 ng/mL. Furthermore, serum spiked experiments showed increment of absorbance with increasing HBsAg concentration, but control trials with other biomolecules showed no increment of absorbance, showing the specific and selective detection of HBsAg. This nanomaterial-modified ELISA plate can detect low levels of HBsAg and help in the diagnosis of HBV infection in its early stages.

Acute and chronic HBV infection in central Argentina: High frequency of sub-genotype F1b, low detection of clinically relevant mutations and first evidence of HDV.

Genomic analysis of hepatitis B virus (HBV) identifies phylogenetic variants, which may lead to distinct biological and clinical behaviors. The satellite hepatitis D virus (HDV) may also influence clinical outcomes in patients with hepatitis B. The aim of this study was to investigate HBV genetic variants, including clinically relevant mutations, and HDV infection in acute and chronic hepatitis B patients in central Argentina. A total of 217 adult HBV infected patients [acute (AHB): n = 79; chronic (CHB): n = 138] were studied; 67 were HBV/human immunodeficiency virus (HIV) coinfected. Clinical and demographic data were obtained from medical records. Serological markers were determined. Molecular detection of HBV and HDV was carried out by RT-Nested PCR, followed by sequencing and phylogenetic analysis. Overall, genotype (gt) F [sub-genotype (sgt) F1b] was the most frequently found. In AHB patients, the gts/sgts found were: F1b (74.7%) > A2 (13.9%) > F4 (7.6%) > C (2.5%) > A1 (1.3%). Among CHB patients: F1b (39.1%) > A2 (23.9%) > F4 (18.2%) > D (9.4%) > C and F6 (3.6% each) > A1, A3 and B2 (0.7% each). The distribution of sgt A2 and gt D was significantly different between HBV mono and HBV/HIV coinfected patients [A2: 15.9% vs. 35.7% (p < 0.05), respectively and D: 14.6% vs. 1.8% (p < 0.05), respectively]. Mutation frequency in basal core promoter/pre-Core (BCP/pC) region was 35.5% (77/217) [AHB: 20.3% (16/79), CHB: 44.2% (61/138)]. In the open reading frame (ORF) S, mutations associated with vaccine escape and diagnostic failure were detected in 7.8% of the sequences (17/217) [AHB: 3.8% (3/79), CHB: 10.1% (14/138)]. ORF-P amino acid substitutions associated with antiviral resistance were detected in 3.2% of the samples (7/217) [AHB: 1.3% (1/79), CHB 4.3%, (6/138)]. The anti-HDV seropositivity was 5.2% (4/77); one sample could be sequenced, belonging to gt HDV-1 associated with sgt HBV-D3. We detected an increase in the circulation of genotype F in Central Argentina, particularly among AHB patients, suggesting transmission advantages over the other genotypes. A low rate of mutations was detected, especially those with antiviral resistance implications, which is an encouraging result. The evidence of HDV circulation in our region, reported for the first time, alerts the health system for its search and diagnosis.

Comparative Study between ELISA and TrueNat for Hepatitis B Virus and Hepatitis C Virus among Antenatal Women Attending Tertiary Care Medical Institute in Manipur, India

Introduction: Viral hepatitis is one of the leading causes of death and disability. Nearly 1.5 million people die every year from Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) related liver diseases. Reported consequences of HBV and HCV infection in pregnancy include an increased likelihood of occurrence of preterm delivery and low birth weight and perinatal transmission. Aim: To compare Enzyme-Linked Immunosorbent Assay (ELISA) and closed system Polymerase Chain Reaction (PCR) TrueNat for the detection of HBV and HCV among antenatal women attending a tertiary care medical institute. Materials and Methods: This was a hospital-based crosssectional study conducted at the Department of Microbiology, JNIMS, Imphal, Manipur, India, during the period from December 2020 to January 2022. Serum samples were tested using ELISA as well as on the TrueNat PCR. Data was analysed using Statistical Package for the Social Sciences (SPSS) version 23.0. Results: Out of 60 samples, using ELISA (QUALISA), 24 were found to be positive for HBV surface antigen (HBsAg) and 14 were found to be positive for HCV antibodies (HCV Ab). On further testing using TrueNat chip-based closed system Real Time-PCR (RT-PCR) Test, 12 were found to be positive for HBV Deoxyribonucleic Acid (DNA) and five were found to be positive for HCV Ribonucleic Acid (RNA). Conclusion: TrueNat PCR could eliminate the problem of false positivity in the detection of HBV and HCV. It is useful to make clinical decisions on starting antiviral therapy and also in documenting the efficacy of the antiviral therapy.

A chemiluminescence based approach to nucleic acid testing to detect HBV, HCV, and HIV-1 in blood screening

Hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) are life-threatening blood-borne infections that can be unintentionally transmitted in transfused blood products. Here, we optimized a method for detecting HBV, HCV, and HIV-1 in blood screening using magnetic nanoparticles (MNPs) and polymerase chain reaction (PCR)-chemiluminescence. Viral genomes were extracted from donated blood samples using MNPs. The isolated viral DNA and RNA were amplified in a one-step parallel reverse transcription-PCR (RT-PCR). HBV, HCV, and HIV-1 were detected using complementary nucleic acid probes and quantified using a chemiluminescent substrate. 10 422 donated blood samples were tested both with the above method and Roche Cobas TaqScreen MPX Test. The lengths of the amplified PCR products for HBV, HCV, and HIV-1 were 119 bp, 220 bp, and 174 bp, respectively, indicating that the extraction methods successfully separated high-quality viral genomes from the serum samples. The probes and reaction conditions used for the chemiluminescent detection of HBV, HCV, and HIV-1 genomes in unknown samples were empirically optimized. Of the 10 422 blood samples screened, 12 were HBV positive and 2 were HCV positive. These results were consistent with those of a parallel-controlled study using Roche Cobas TaqScreen MPX Test. The assay described here is fast, accurate, and sensitive for detecting HBV, HCV, and HIV, which runs in parallel and has broad implications for blood screening and epidemiological studies.

Molecular detection of HHV-1, HHV-2, HHV-5 and HBV in semen of fertile and infertile men by multiplex PCR method.

A great diversity of factors including viruses such as human herpes virus 1&2 (HHV-1&2), human herpes virus 5 (HHV-5), and hepatitis B virus (HBV) play key roles in sterility and it is worth noting that male infertility accounts for nearly 50% of barrenness, globally. In this regard, we evaluated the prevalence of the aforementioned viruses in semen specimens of two distinct groups of men referred to Novin Infertility Center in Mashhad, Iran. In this cross-sectional study, 300 semen samples were collected from 150 infertile and 150 fertile men. Subsequently, genomic DNA was extracted before performing multiplex polymerase chain reaction (PCR). Eventually, the results were analyzed via SPSS Statistics V.16.0. Out of 300 specimens, 183 (61.1%) were positive at least for one of the forenamed viruses; genome detection of HHV-1&2, HHV-5, and HBV were 27%, 18%, 36.66%, and 4%, respectively. The current study found no correlation between infertility and HBV, HHV-5, and HHV-1&2, which may have to do with factors like sample size, the geographical distribution of the viruses, and the lifestyle (sexual behavior) of the participants. These results emphasize the implementation of such studies on a broader scale to determine the exact factors involved in infertility.

False occult hepatitis B virus infection with negative HBsAg while positive HBeAg &amp; overt DNA levels: Case report and literature review

Background: Occult Hepatitis B Infection (OBI) is characterized by the detection of Hepatitis B Virus (HBV) DNA in serum (usually HBV DNA &lt; 200 IU/ml) or liver but negativity for hepatitis B surface antigen. The diagnosis of OBI is based on the sensitivity of assays used in the detection of HBV DNA and HBsAg. HBsAg assays with inadequate sensitivity or inability to detect HBV S variants may lead to a false negative HBsAg result and misdiagnosis of OBI in people with overt HBV infection. An OBI patient was followed 4 years with the analysis of HBV detection.