• Component Description
• Eligible Sample
• Description of Laboratory Methodology
• Laboratory Method Files
• Laboratory Quality Assurance and Monitoring
• Data Processing and Editing
• Analytic Notes
• References
• Codebook
  • SEQN - Respondent sequence number
  • LBD9M6N - HPV 6 (IU/mL)
  • LBD96NLC - HPV 6 (Quantitative) comment code
  • LBD9M11N - HPV 11 (IU/mL)
  • LBD911LC - HPV 11 (Quantitative) comment code
  • LBD9M16N - HPV 16 (IU/mL)
  • LBD916LC - HPV 16 (Quantitative) comment code
  • LBD9M18N - HPV 18 (IU/mL)
  • LBD918LC - HPV 18 (Quantitative) comment code
  • LBD9M31N - HPV 31 (IU/mL)
  • LBD931LC - HPV 31 (Quantitative) comment code
  • LBD9M33N - HPV 33 (IU/mL)
  • LBD933LC - HPV 33 (Quantitative) comment code
  • LBD9M45N - HPV 45 (IU/mL)
  • LBD945LC - HPV 45 (Quantitative) comment code
  • LBD9M52N - HPV 52 (IU/mL)
  • LBD952LC - HPV 52 (Quantitative) comment code
  • LBD9M58N - HPV 58 (IU/mL)
  • LBD958LC - HPV 58 (Quantitative) comment code
  • LBX9M6 - HPV6 9-plex coded result
  • LBX9M11 - HPV11 9-plex coded result
  • LBX9M16 - HPV16 9-plex coded result
  • LBX9M18 - HPV18 9-plex coded result
  • LBX9M31 - HPV31 9-plex coded result
  • LBX9M33 - HPV33 9-plex coded result
  • LBX9M45 - HPV45 9-plex coded result
  • LBX9M52 - HPV52 9-plex coded result
  • LBX9M58 - HPV58 9-plex coded result

National Health and Nutrition Examination Survey

2015-2016 Data Documentation, Codebook, and Frequencies

Human Papillomavirus (HPV) - 6, 11, 16, 18, 31, 33, 45, 52, & 58 Antibody – Serum: M9ELISA (HPVN_I_R)

RDC Only Data File: HPVN_I_R.xpt

First Published: September 2024

Last Revised: NA

Due to disclosure issues, this dataset is not available publicly. Access may be obtained through the NCHS Research Data Center (https://www.cdc.gov/rdc).

Component Description

Human papillomavirus (HPV) infection is the most common sexually transmitted infection in the United States, and persistent infection with certain types of HPV can progress to cancer. HPV-attributable cancers include cervical, vaginal, vulvar, anal, penile, and oropharyngeal cancers. NHANES offers a unique opportunity to assess the prevalence of HPV infection in the general population and the impact of HPV vaccination.

HPV vaccination was introduced in the national immunization program for females in 2006 and for males in 2011. Reducing infections of HPV types prevented by the vaccine in young adults and increasing the proportion of adolescents vaccinated for HPV are both Healthy People 2030 objectives. Knowledge of the national prevalence of HPV infection is critical for planning vaccination strategies and monitoring the impact of vaccination in the United States. Seroprevalence reflects current and past exposure to HPV as well as presence of vaccine-induced antibody and can provide important information about HPV epidemiology and vaccination.

Eligible Sample

Examined participants aged 14 to 59 years were eligible.

Description of Laboratory Methodology

M9ELISA is an assay developed for the detection of 9 HPV types, which includes both qualitative and quantitative results.

M9ELISA allows for simultaneous detection of antibodies to HPV 6, 11, 16, 18, 31, 33, 45, 52, and 58 in one microwell (Panicker et. al., 2021). Virus-like particles (VLP), self-assembled HPV L1+L2 protein capsids that resemble intact virions, are used as antigen in the assay. HPV-type specific antibodies present in test samples will bind to type-specific conformational epitopes on the VLPs adsorbed onto the surface of the microwell. Bound human IgG is detected with a SulfoTAG^TM  Anti-Human IgG using electrochemiluminescent detection on the Meso Scale Discovery (MSD) plate reader that generates relative light units (RLU). The amount of antibodies in the sample is estimated relative to a reference sample using the parallel line method and reported in International Units/milliliter (IU/mL). Samples are scored as positive or negative using a pre-determined “cut-off” level. The assay is used for research purposes and does not have clinical relevance.  

Refer to the Laboratory Method Files section for a detailed description of the laboratory methods used.

Laboratory Method Files

M9ELISA – Multiplexed VLP-based IgG ELISA on the Meso Scale Discovery Platform Lab Procedure Manual (September 2024)

Laboratory Quality Assurance and Monitoring

Serum samples were processed, stored, and shipped to the Chronic Viral Diseases Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA for analysis.

Detailed instructions on specimen collection and processing are discussed in the NHANES Laboratory Procedures Manual (LPM). Vials were stored under appropriate frozen conditions (-20°C) until they were shipped to the National Center for Emerging and Zoonotic Infectious Diseases for testing. The M9ELISA was conducted in the HPV Immunology lab at the Centers for Disease Control and Prevention.

The NHANES quality assurance and quality control (QA/QC) protocols meet the 1988 Clinical Laboratory Improvement Amendments mandates. Detailed QA/QC instructions are discussed in the NHANES LPM.

Mobile Examination Centers (MECs)

Laboratory team performance is monitored using several techniques. NCHS and contract consultants use a structured competency assessment evaluation during visits to evaluate both the quality of the laboratory work and the QC procedures. Each laboratory staff member is observed for equipment operation, specimen collection and preparation; testing procedures and constructive feedback are given to each staff member. Formal retraining sessions are conducted annually to ensure that required skill levels were maintained.

Analytical Laboratories

NHANES uses several methods to monitor the quality of the analyses performed by the contract laboratories. In the MEC, these methods include performing blind split samples collected on “dry run” sessions. In addition, contract laboratories randomly perform repeat testing on 2% of all specimens.

NCHS developed and distributed a quality control protocol for all CDC and contract laboratories, which outlined the use of Westgard rules (Westgard et. al., 1981) when running NHANES specimens. Progress reports containing any problems encountered during shipping or receipt of specimens, summary statistics for each control pool, QC graphs, instrument calibration, reagents, and any special considerations are submitted to NCHS quarterly. The reports are reviewed for trends or shifts in the data. The laboratories are required to explain any identified areas of concern.

Data Processing and Editing

The data were reviewed. Incomplete data or improbable values were sent to the performing laboratory for confirmation.

Analytic Notes

Refer to the 2015-2016 Laboratory Data Overview for general information on NHANES laboratory data.

There are over 800 laboratory tests performed on NHANES participants. However, not all participants provided biospecimens or enough volume for all the tests to be performed. The specimen availability can also vary by age or other population characteristics. Analysts should evaluate the extent of missing data in the dataset related to the outcome of interest as well as any predictor variables used in the analyses to determine whether additional re-weighting for item non-response is necessary.

Please refer to the NHANES Analytic Guidelines and the on-line NHANES Tutorial for details on the use of sample weights and other analytic issues.

Sample Weights

MEC exam sample weights should be used for analyses.

Demographic and Other Related Variables

The analysis of NHANES 2015-2016 laboratory data must be conducted using the appropriate survey design and demographic variables. The NHANES 2015-2016 Demographics File contains demographic data, health indicators, and other related information collected during household interviews as well as the sample design variables. The recommended procedure for variance estimation requires use of stratum and PSU variables (SDMVSTRA and SDMVPSU, respectively) in the demographic data file.

This laboratory data file can be linked to the other NHANES data files using the unique survey participant identifier (i.e., SEQN).

Detection Limits

Two variables are provided for each of these analytes. The variable name ending in “LC” (ex., LBD9M6LC) indicates whether the result was below the limit of quantitation: the value “0” means that the result was at or above the limit of quantitation, “1” indicates that the result was below the limit of quantitation. For analytes with analytic results below the lower limit of quantitation (ex. LBD9M6LC =1), an imputed fill value was placed in the analyte results field. This value is the lower limit of quantitation divided by the square root of 2 (LLOQ/sqrt[2]). The other variable prefixed “LBD” and ended with an “N” (ex., LBD9M6N) provides the analytic result for the analyte.

The lower limit of quantitation (LLOQ, in units) for HPV antibody 6, 11, 16, 18, 31, 33, 45, 52, & 58 are:

VARIABLE	ANALYTE NAME	LLOQ
LBD9M6N	HPV 6 (Quantitative)	2.2 IU/ml
LBD9M11N	HPV 11 (Quantitative)	2.6 IU/ml
LBD9M16N	HPV 16 (Quantitative)	1.2 IU/ml
LBD9M18N	HPV 18 (Quantitative)	0.4 IU/ml
LBD9M31N	HPV 31 (Quantitative)	2.2 IU/ml
LBD9M33N	HPV 33 (Quantitative)	3.6 IU/ml
LBD9M45N	HPV 45 (Quantitative)	1.2 IU/ml
LBD9M52N	HPV 52 (Quantitative)	4.3 IU/ml
LBD9M58N	HPV 58 (Quantitative)	4.0 IU/ml

Note: LLOQ – lower limit of quantitation; IU/ml – International Units/milliliter

Serostatus Cut-off Value

The qualitative results for the analytes are provided in variables prefixed “LBX” (ex., LBX9M6). Serostatus cut-off value for a positive result for each HPV analyte was established as below. An antibody concentration below this value is coded as negative and a value equal or above is considered positive.

VARIABLE	ANALYTE NAME	Cut-off Value
LBX9M6	HPV 6 (Qualitative)	7.6 IU/ml
LBX9M11	HPV 11 (Qualitative)	5.2 IU/ml
LBX9M16	HPV 16 (Qualitative)	1.6 IU/ml
LBX9M18	HPV 18 (Qualitative)	3.5 IU/ml
LBX9M31	HPV 31 (Qualitative)	5.5 IU/ml
LBX9M33	HPV 33 (Qualitative)	6.8 IU/ml
LBX9M45	HPV 45 (Qualitative)	4.2 IU/ml
LBX9M52	HPV 52 (Qualitative)	6.2 IU/ml
LBX9M58	HPV 58 (Qualitative)	12.8 IU/ml

Note: IU/ml – International Units/milliliter

References

• Panicker G, Rajbhandari I, Pathak HN, Brady AM, Unger ER. Multiplex immunoassay to measure antibody response to nine HPV vaccine types. Journal of Immunological Methods 2021. https://doi.org/10.1016/j.jim.2021.113136
• Westgard J.O., Barry P.L., Hunt M.R., Groth T. A multi-rule Shewart chart for quality control in clinical chemistry. Clin Chem (1981) 27:493-501

Codebook and Frequencies