Introduction

Viral hepatitis remains one of the leading causes of infectious disease mortality worldwide. According to the World Health Organization, an estimated 254 million people were living with chronic hepatitis B virus (HBV) infection and 50 million with hepatitis C virus (HCV) infection in 2022. These infections resulted in 1.3 million deaths globally that year — equivalent to tuberculosis mortality — with HBV accounting for 83% and HCV for 17% of hepatitis-related deaths. Every day, approximately 3,500 people die from viral hepatitis complications (1,2).

India bears a disproportionately high burden, with approximately 29–30 million individuals chronically infected with HBV and 5.5 million with HCV, ranking the country second globally in absolute number of cases (3,4). National estimates indicate an overall HBV seroprevalence of around 0.95% and HCV prevalence of 0.32%, with notable regional variations; higher rates are observed in rural areas and tribal populations for HBV, and in certain urban pockets for HCV (5). Transmission continues primarily through unsafe injections, contaminated blood products, high-risk behaviours, and perinatal routes. Although universal childhood HBV vaccination has contributed to declining new infections in younger age groups, significant gaps in infection control, awareness, and service coverage persist (6).

Despite the availability of a safe and effective HBV vaccine and highly curative direct-acting antiviral therapy for HCV, diagnosis and treatment rates in India remain suboptimal. This slow progress has placed the country off-track to achieve the World Health Organization’s 2030 viral hepatitis elimination targets (1). Tertiary care hospitals serve as critical referral centres that capture a wide spectrum of cases, including advanced liver disease, and provide essential real-world data on epidemiological trends, demographic patterns, and co-infections.

Previous hospital-based studies in India have documented varying seroprevalence rates of HBV and HCV, often showing male predominance and higher detection among middle-aged adults. However, contemporary data reflecting post-pandemic healthcare utilisation, evolving risk profiles, and the impact of the National Viral Hepatitis Control Programme (launched in 2018) are limited, particularly from the East Delhi region (7).

The present retrospective study was conducted to evaluate the trends in the epidemiology of HBV and HCV infections among patients attending a tertiary care government hospital in East Delhi over a 2-year period. By analysing seroprevalence rates, demographic characteristics, age- and gender-specific distributions, this study aims to provide updated insights into the local disease burden. Such information is vital for strengthening targeted screening strategies, optimising resource allocation, and supporting evidence-based interventions under the National Viral Hepatitis Control Programme to accelerate progress toward viral hepatitis elimination in the region.

Materials and Methods

This retrospective observational study was conducted at the Virology Laboratory, Department of Microbiology, University College of Medical Sciences and Guru Teg Bahadur Hospital, Delhi. Laboratory records over a two-year period, from 1st January 2024 to 31st December 2025, were systematically reviewed.

The study population comprised patients attending outpatient services or admitted to the hospital who underwent serological testing either as part of routine government-mandated screening or for diagnostic evaluation.

Data pertaining to enzyme-linked immunoassay results for Hepatitis B surface antigen (HBsAg) and Hepatitis C virus (HCV) antigen and/or anti-HCV antibodies were extracted and analysed. Serological screening for hepatitis B surface antigen (HBsAg) and anti-hepatitis C virus (anti-HCV) antibodies was performed using commercially available enzyme-linked immunosorbent assay (ELISA) kits—HEPALISA (HBsAg) and MICROLISA anti-HCV (J. Mitra & Co. Pvt. Ltd., India)—in accordance with the manufacturer’s instructions.

Both assays are third generation Elisa, based on the solid-phase ELISA principle employing microwells coated with specific monoclonal antibodies or antigens. Serum or plasma samples were added to the wells and incubated with enzyme-labelled conjugates, facilitating the formation of antigen–antibody complexes in reactive samples. Following washing steps to remove unbound material, a chromogenic substrate was added, resulting in a colorimetric reaction proportional to the presence of target analyte. The optical density (OD) was measured at 450 nm using an ELISA reader. Results were interpreted as reactive or non-reactive based on the assay-specific cut-off values as defined by the manufacturer.

For screening purposes, the detection of HBsAg and anti-HCV antibodies was performed using the ELISA method, the Patient data were extracted from the system in Excel format. The following information was collected for each patient: age, sex, HBsAg, and anti-HCV Ab ELISA results. The analysis was conducted exclusively on data provided for routine laboratory investigations. No additional data were extracted, and only variables documented as part of standard diagnostic and reporting processes were included in the study. Descriptive statistics was utilized to summarize the study population characteristics. Statistical analyses were done with the SPSS 27.0 software, and p-values of <0.05 were considered statistically significant.

Results

During the study period, a total of 63,616 samples were screened for Hepatitis B and 50,951 samples for Hepatitis C. Of these, 1,468 samples tested positive for Hepatitis B (2.31%), while 2,426 samples were positive for Hepatitis C (4.76%).

The overall HBsAg positivity remained stable over the two-year period, with 2.319% in 2024 and 2.296% in 2025 (Table 1). In contrast, Anti-HCV seropositivity showed a slight increase from 4.526% in 2024 to 5.078% in 2025 (Table 2).

The monthly distribution of cases demonstrated distinct temporal patterns (Figure 1). Hepatitis B cases showed moderate fluctuations across both years, with a peak in October 2024 (97 cases) and a secondary rise in December 2025 (91 cases). In contrast, Hepatitis C exhibited greater variability, with a pronounced peak in May of both years (183 and 184 cases), along with elevated numbers during June–July, followed by a marked decline in the latter half of 2025.

Gender-wise analysis revealed a marked male predominance in both infections (Figure 2). Among Hepatitis B cases, 1001 (68.2%) were males and 467 (31.8%) were females (male: female = 2:1). In Hepatitis C, 1886 (77.7%) cases were males and 540 (22.3%) were females (male: female = 3:1). The difference in gender distribution between the two infections was statistically highly significant (p < 0.00001).

Age-stratified analysis demonstrated that the majority of cases were observed in younger age groups (Figure 3). The 21–40 years age group accounted for the highest proportion, followed by the 41–60 years group, while paediatric (0–20 years) and elderly (≥61 years) populations contributed relatively smaller proportions.

Analysis based on indication for testing showed differing patterns between the two infections (Figure 4). Given the paucity of clinical background data available with the laboratory, a presumptive grouping was attempted to understand the demand for testing from different clinical departments. Therefore, the authors grouped the dataset with the following groups.

Group 1, comprising symptomatic patients from the Department of Medicine; Group 2, antenatal screening cases from the ANC clinic; Group 3, individuals undergoing pre-operative screening across surgical specialties, including General Surgery, Orthopaedics, and ENT; and Group 4, patients attending the ART clinic. This systematic stratification reflects the diverse clinical settings in which hepatitis screening is undertaken and enables a more robust and nuanced interpretation of disease distribution across heterogeneous patient populations.

Among hepatitis B cases, the highest proportion was observed among symptomatic patients referred from the Department of Medicine, followed by individuals attending the ART clinic and those undergoing pre-operative screening, with a minimal contribution from antenatal screening. In contrast, hepatitis C cases were predominantly identified among ART clinic attendees, followed by symptomatic patients and individuals screened pre-operatively across various surgical specialties. Notably, the majority of these cases were derived from outpatient services, reflecting the predominance of routine screening and follow-up evaluations in the outpatient setting.

Figure 1: Monthly distribution of Hepatitis B (solid lines) and Hepatitis C (dotted lines) positive cases in 2024 and 2025.

Figure 2: Comparison of gender distribution in Hepatitis B and C cases (n).

Figure 3: Age-stratified Trends in Hepatitis B and Hepatitis C cases (n)

Figure 4: Comparative Distribution of Hepatitis B and Hepatitis C by Indication for Testing (%)

Discussion

The present study provides a comprehensive evaluation of the Sero-epidemiological trends of Hepatitis B and Hepatitis C infections in a tertiary care hospital setting in East Delhi. The large sample size and inclusion of both outpatient and inpatient populations strengthen the reliability of the findings and reflect real-world disease burden in an urban North Indian healthcare setting.

The overall seroprevalence of Hepatitis B (2.31%) observed in this study falls within the intermediate endemicity range reported for India (2–4%) (8,9). This finding is consistent with several hospital-based and community-based studies conducted across the country, indicating a relatively stable burden of HBV infection. In contrast, the seropositivity of Hepatitis C (4.76%) is notably higher than national estimates, which range between 0.3% and 1% (10,11). This discrepancy may be attributed to the tertiary care nature of the study setting, where a higher proportion of high-risk individuals, including those attending ART clinics and those undergoing diagnostic workups, are represented.

A key observation of this study is the stable trend in Hepatitis B positivity over the two-year period. This plateau likely reflects the cumulative impact of public health interventions, including universal immunization, improved blood transfusion safety, and increased awareness (12,13). Similar stabilization trends have been documented in recent Indian studies, particularly among younger populations benefiting from vaccination programs. However, the persistence of HBV infection highlights ongoing transmission in unvaccinated adults and the need for expanded screening and immunization strategies.

In contrast, Hepatitis C demonstrated a relatively higher and slightly increasing trend, suggesting ongoing transmission dynamics. This is in agreement with recent studies that emphasize the role of unsafe injections, invasive procedures, and high-risk behaviours in sustaining HCV transmission in developing countries (14,15). The predominance of HCV cases among ART clinic attendees in the present study further supports the established association between HCV infection and high-risk populations, including individuals with HIV infection and people who inject drugs.

Temporal analysis in this study did not reveal a consistent seasonal pattern for Hepatitis B, whereas Hepatitis C showed clustering during mid-year months. Such variations are unlikely to represent true seasonality and may instead reflect fluctuations in healthcare utilization, targeted screening activities, or episodic exposure risks (16).

The male predominance observed in both infections, particularly in Hepatitis C (male: female ratio of 3:1), is consistent with previous Indian and global studies (14,17). This pattern may be explained by increased exposure to occupational hazards, higher likelihood of high-risk behaviours, and gender-related differences in healthcare-seeking behaviour.

Age-wise distribution revealed that the 21–40 years age group accounted for the highest proportion of cases, followed by the 41–60 years group. This trend is consistent with existing epidemiological evidence and reflects increased exposure to risk factors during the most active years of life (18,19). The relatively lower burden in paediatric populations supports the success of hepatitis B vaccination programs, whereas the lower proportion in elderly individuals may reflect under-screening or survivor bias.

An important finding of this study is the high proportion of asymptomatic cases, particularly for Hepatitis B, indicating that a substantial number of infections remain undiagnosed until detected incidentally. This observation aligns with previous studies emphasizing the silent nature of viral hepatitis and the importance of opportunistic screening (20,21).

The higher contribution of cases from departments such as Medicine and ART clinics further highlights the importance of integrated screening strategies within hospital settings, especially for high-risk populations. Opportunistic testing in such settings plays a critical role in early diagnosis and prevention of disease progression.

Recognizing the critical role of surveillance in achieving hepatitis elimination, the Government of India has placed strong emphasis on strengthening surveillance systems under the National Viral Hepatitis Control Program, launched on 28 July 2018 in alignment with World Health Organization targets for 2030 (20). Implemented under the Ministry of Health and Family Welfare, the program provides free screening, diagnosis, and treatment, supported by a nationwide network of laboratories enabling standardized testing, systematic data collection, and robust reporting (22). Despite these efforts, challenges such as low diagnosis rates, limited access to advanced diagnostics, regional disparities, and suboptimal birth-dose vaccination continue to impede progress toward elimination goals (23).

In conclusion, the study demonstrates a stable endemic pattern of Hepatitis B alongside a relatively higher and evolving burden of Hepatitis C, particularly among high-risk populations. These findings underscore the need for strengthening targeted screening programs, improving access to antiviral therapy, and reinforcing infection control measures. Continuous surveillance and region-specific epidemiological data are essential to guide public health strategies and achieve the goal of viral hepatitis elimination.

Despite its strengths, this study has certain limitations. Being retrospective and hospital-based, the findings may not be fully generalizable to the broader community. Additionally, lack of detailed risk factor assessment and follow-up data limits causal inference and long-term outcome analysis.

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