Hepatitis viruses constitute a major public health problem because of the morbidity and mortality associated with the acute and chronic consequences of these infections. Co-infection with hepatitis D virus (HDV) in persons with acute or chronic hepatitis B virus (HBV) infection can lead to fulminant hepatitis. New immunization strategies have been developed to eliminate the spread of HBV and hepatitis A virus (HAV) in the United States. Recommendations have also been developed for the prevention and control of hepatitis C virus (HCV) infection. Because of the high rate of asymptomatic infection with these viruses, information about the prevalence of these diseases is needed to monitor prevention efforts. By testing a nationally representative sample of the U.S. population, NHANES will provide the most reliable estimates of age-specific prevalence needed to evaluate the effectiveness of the strategies to prevent these infections. In addition, NHANES provides the means to better define the epidemiology of other hepatitis viruses. NHANES testing for markers of infection with hepatitis viruses will be used to determine secular trends in infection rates across most age and racial/ethnic groups, and will provide a national picture of the epidemiologic determinants of these infections.
All participants aged 6 years or older are eligible to be tested.
Serum specimens are processed, stored, and shipped to the Division of Viral Hepatitis, National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention, Centers for Disease Control and Prevention. Detailed specimen collection and processing instructions are discussed in the NHANES Laboratory/Medical Technologists Procedures Manual (LPM).
Hepatitis B core antibody (anti-HBc)
The VITROS Anti-HBc assay is performed using the VITROS Anti-HBc Reagent Pack and VITROS Immunodiagnostic Products Anti-HBc Calibrator on the VITROS ECi/ECiQ or VITROS 3600 Immunodiagnostic System.
A competitive immunoassay technique is used. This involves the reaction of anti-HBc in the sample with hepatitis B core antigen (HBcAg) coated wells. Unbound sample is removed by washing. Horseradish peroxidase (HRP)-labeled antibody conjugate (mouse monoclonal anti-HBc) is then allowed to react with the remaining exposed HBcAg on the well surface. Unbound conjugate is removed by washing.
The bound HRP conjugate is measured by a luminescent reaction. A reagent containing luminogenic substrates (a luminol derivative and a peracid salt) and an electron transfer agent is added to the wells. The HRP in the bound conjugate catalyzes the oxidation of the luminol derivative, producing light. The electron transfer agent increases the level and duration of the light produced. The light signals are read by the VITROS ECi/ECiQ or VITROS 3600 Immunodiagnostic System. The amount of HRP conjugate bound is indicative of the concentration of anti-HBc present in the sample.
Hepatitis B surface antigen (HBsAg)
The VITROS HBsAg assay is performed using the VITROS HBsAg Reagent Pack and VITROS Immunodiagnostic Products HBsAg Calibrator on the VITROS ECi/ECiQ or VITROS 3600 Immunodiagnostic System.
An immunometric technique is used. This involves the simultaneous reaction of HBsAg in the sample with mouse monoclonal anti-HBs antibody coated onto the wells and a horseradish peroxidase (HRP)-labeled mouse monoclonal anti-HBs antibody in the conjugate. Unbound conjugate is removed by washing.
A reagent containing luminogenic substrates (a luminol derivative and a peracid salt) and an electron transfer agent is added to the wells. The HRP in the bound conjugate catalyzes the oxidation of the luminol derivative, producing light. The electron transfer agent increases the level and duration of the light produced. The light signals are read by the VITROS ECi/ECiQ or VITROS 3600 Immunodiagnostic System. The amount of HRP conjugate bound is indicative of the level of HBsAg present in the sample.
Hepatitis D antibody (anti-HDV)
The DiaSorin ETI-AB-DELTAK-2 enzyme immunoassay used for qualitative anti-HDV determination is a simultaneous competitive assay. Anti-HDV present in the sample and labeled anti-HDV antibodies compete for a fixed quantity of HDAg bound to the solid phase. The quantity of enzyme tracer bound to the solid phase and consequently the enzyme activity are inversely proportional to the anti-HDV concentration present in samples or controls.
Enzyme activity is measured by adding a colorless chromogen/substrate solution. The enzyme action on the chromogen/substrate produces a color which is measured with a photometer.
The analytical methods are described in the Description of Laboratory Methodology section above.
The NHANES quality assurance and quality control (QA/QC) protocols meet the 1988 Clinical Laboratory Improvement Amendments mandates. Detailed quality control and quality assurance instructions are discussed in the NHANES Laboratory/Medical Technologists Procedures Manual (LPM).
A detailed description of the quality assurance and quality control procedures can be found on the NHANES website.
Refer to the 2011-2012 Laboratory Data Overview for general information on NHANES laboratory data.
The analysis of NHANES 2011-2012 laboratory data must be conducted using the appropriate survey design and demographic variables. The NHANES 2011-2012 Demographics File contains demographic data, health indicators, and other related information collected during household interviews as well as the sample weight variables. The Fasting Questionnaire File includes auxiliary information such as fasting status, the time of venipuncture, and the conditions precluding venipuncture. The demographics and fasting questionnaire files may be linked to the laboratory data file using the unique survey participant identifier (i.e., SEQN).
The age range and constraints for hepatitis B and D testing are as follows:
Hep B
The hepatitis B core antibody test is performed on all examinees 6 years old or older while the hepatitis B surface antibody test is performed on all examinees 2 years old or older. The Hepatitis B surface antigen is tested only when the Hepatitis B core antibody test is positive. Participant results are coded positive for surface antigen if the surface antigen test is positive; they are coded negative for surface antigen if the test for surface antigen is negative or if the test for hepatitis B core antibody is negative.
Hep D
The Hepatitis Delta Virus (HDV) is a RNA defective virus and infection with HDV only occurs in the presence of acute or chronic HBV infection. In NHANES, the test for antibody to HDV is performed on participants 6 years of age or older who test positive for anti-HBc and HBsAg. The denominator for anti-HDV includes all anti-HBc negative samples, the anti-HBc positive samples that were subsequently found to be HBsAg negative, and the samples that were anti-HBc positive and HBsAg positive that were subsequently found to be anti-HDV negative.
Exam sample weights should be used for analyses. Please refer to the NHANES Analytic Guidelines and the on-line NHANES Tutorial for further details on the use of sample weights and other analytic issues.
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 468 | 468 | |
| 2 | Negative | 6598 | 7066 | |
| . | Missing | 755 | 7821 |
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 43 | 43 | |
| 2 | Negative | 7022 | 7065 | |
| . | Missing | 756 | 7821 |
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 5 | 5 | |
| 2 | Negative | 7060 | 7065 | |
| . | Missing | 756 | 7821 |