Hepatitis viruses constitute a major public health problem because of the morbidity and mortality associated with the acute and chronic consequences of these infections. New immunization strategies have been developed to eliminate the spread of hepatitis B virus (HBV) 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.
Hepatitis B: core antibody and surface antigen; Hepatitis C: confirmed antibody and RNA (HCV-RNA); Hepatitis D antibody: participants aged 6 years and older are eligible to be tested.
Hepatitis B Virus Core Antibody
The Ortho HBc ELISA Test System is a qualitative enzyme-linked immunosorbent assay (ELISA) for the detection of total antibody to anti-HBc in human serum or plasma. Anti-HBc appears in virtually all individuals infected with HBV and is an accurate serological marker of current and past infection.
ELISA procedures provide a means for routinely detecting antibodies to specific antigens. This FDA-licensed method is commercially obtained in kit form. The literature and instructions with each kit constitute the standard operating procedure (SOP) for the method.
Hepatitis B Surface Antigen
The AUSZYME Monoclonal test is a solid-phase “sandwich” enzyme immunoassays used to detect the presence of HBsAg, which indicates current infection with HBV. Specimens that test nonreactive by the AUSZYME Monoclonal tests (Abbott Diagnostics) are considered negative for HBsAg and are not tested further. All specimens considered reactive initially are repeat-tested in duplicate using the same procedure as that used in the initial test. If neither of the repeat tests is reactive, the specimen is considered negative for HBsAg. If the specimen is reactive in either of the repeat tests, the sample is considered repeatedly reactive.
Hepatitis C Antibody
Qualitative determination of the human antibody directed against hepatitis C virus (anti-HCV) in human serum or plasma is measured using direct solid-phase enzyme immunoassay with the anti-HCV screening ELISA. Positive specimens are repeated in duplicate according to the same procedure. Repeatedly positive specimens are tested supplementally using the Chiron RIBA Processor System (Chiron Corporation, Inc.). Samples where the RIBA result was positive are reported as confirmed positive for antibody to HCV Samples where the RIBA result was negative are reported as negative for antibody to HCV and indeterminate results are reported as indeterminate.
The Chiron RIBA HCV 3.0 Strip (confirmation test)
The Chiron RIBA 3.0 Strip Immunoblot Assay (SIA; Chiron Corporation, Inc.) is an in vitro qualitative enzyme immunoassay for the detection of antibody to hepatitis C virus (anti-HCV) in human serum or plasma.
Detection of anti-HCV by SIA methodology is based upon traditional Western and dot blotting techniques, in which specific immunogens (i.e. antigenic polyproteins) encoded by the HCV genome are immobilized onto a membrane support. Visualization of anti-HCV reactivity in specimens to the individual HCV-encoded proteins is accomplished with anti-human IgG enzyme-conjugates in conjunction with a colorimetric enzyme substrate. Qualitative determination of the human antibody directed against hepatitis C virus (anti-HCV) in human serum or plasma is measured with a direct solid-phase enzyme immunoassay. A detailed description of the laboratory method used can be found on the NHANES website.
Hepatitis C RNA (HCV-RNA)
The COBAS AMPLICOR HCV MONITOR Test, version 2 0 (v2.0) is an in vitro nucleic acid amplification test for the quantitation of Hepatitis C Virus RNA in human serum or plasma on the COBAS AMPLICOR Analyzer.
Hepatitis D Antibody
The International Immunodiagnostics HDV Ab assay is a competitive enzyme immunoassay (ELISA) for the determination of antibodies to Hepatitis D Virus or HDV in human plasma and sera with a 'two steps" methodology. Anti-HDV antibodies, if present in the sample, compete with a virus specific polyclonal IgG, labeled with peroxidase (HRP), for a fixed amount of rec-HDV coated on the Microplate, in "two step" incubation. The concentration of the bound enzyme on the solid phase is inversely proportional to the amount of anti-HDV antibodies in the sample and its activity is detected by adding the chromagen/substrate in the second incubation. The concentration of HDV-specific antibodies in the sample is determined by means of a cut-off value that allows for the semi-quantitative detection of anti-HDV antibodies.
Blood specimens are processed, stored, and shipped to the Division of Viral Hepatitis, National Center for Infectious Diseases, National Centers for Disease Control and Prevention. Detailed specimen collection and processing instructions are discussed in the NHANES LPM. Read the LABDOC file for detailed data processing and editing protocols. The analytical methods are described in the Analytic Notes for Data Users section below. Detailed instructions on specimen collection and processing can be found on the NHANES website.
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). Read the LABDOC file for detailed QA/QC protocols. A detailed description of the quality assurance and quality control procedures can be found on the NHANES website.
The analysis of NHANES laboratory data must be conducted with the key survey design and basic demographic variables. The NHANES Household Questionnaire Data Files contain demographic data, health indicators, and other related information collected during household interviews. They also contain all survey design variables and sample weights for these age groups. The phlebotomy file includes auxiliary information such as the conditions precluding venipuncture. The household questionnaire and phlebotomy files may be linked to the laboratory data file using the unique survey participant identifier SEQN.
The age ranges and constraints for hepatitis testing are as follows:
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 339 | 339 | |
| 2 | Negative | 6782 | 7121 | |
| . | Missing | 1466 | 8587 |
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 17 | 17 | |
| 2 | Negative | 7104 | 7121 | |
| . | Missing | 1466 | 8587 |
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 76 | 76 | |
| 2 | Negative | 7036 | 7112 | |
| 5 | Indeterminate | 16 | 7128 | |
| . | Missing | 1459 | 8587 |
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 35 | 35 | |
| 2 | Negative | 21 | 56 | |
| . | Missing | 8531 | 8587 |
| Code or Value | Value Description | Count | Cumulative | Skip to Item |
|---|---|---|---|---|
| 1 | Positive | 3 | 3 | |
| 2 | Negative | 7118 | 7121 | |
| . | Missing | 1466 | 8587 |