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Protocol - Adipocytokines

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Description

This protocol provides instructions for drawing, processing, and storing blood according to the National Health and Nutrition Examination Survey (NHANES). The bioassay of leptin and high molecular weight adiponectin uses commercial ELISA kits.

Specific Instructions

To aid in comparability, the Obesity Working Group recommends that the investigator record the make and manufacturer of equipment used and the repeatability and coefficients of variation for the assay.

Availability

Available

Protocol

The following is a summary version of the full National Health and Nutrition Examination Survey (NHANES) 2007-2008 protocol.

Exclusion Criteria

Persons will be excluded from this component if they:

  • report that they have hemophilia, or
  • report that they have received cancer chemotherapy in the last 4 weeks.

SP = Sample Person

1. Do you have hemophilia?

1 [ ] Yes

2 [ ] No

7 [ ] Refused

9 [ ] Don’t Know

If the SP answers "Yes," the SP is excluded from the blood draw.

If the SP answers "No" or "Don’t Know," blood is drawn from the SP.

2. Have you received cancer chemotherapy in the past 4 weeks, or do you anticipate such therapy in the next 4 weeks?

1 [ ] Yes

2 [ ] No

7 [ ] Refused

9 [ ] Don’t Know

If the SP answers "Yes," the SP is excluded from the blood draw.

If the SP answers "No" or "Don’t Know," blood is drawn from the SP.

Note from the Obesity/Adiposity Working Group (WG): The investigator should record the reason a person is excluded from the blood draw.

Venipuncture Procedures

Editor’s Note: Please review chapter 4 of the Laboratory Procedures Manual from the NHANES for a full description of phlebotomy procedures: 2007-2008 NHANES Lab Manual.

Venipuncture should generally be performed using the median cubital, cephalic, or basilica veins in the left arm unless this arm is unsuitable. If the veins in the left arm are unsuitable, look for suitable veins on the right arm. If the veins in the antecubital space on both arms are not suitable, then look for veins in the forearm or dorsal side of the hand on the left arm/hand and then right arm/hand.

Recording the Results of the Venipuncture Procedure

Immediately after completing the venipuncture, record the results of the blood draw, the reasons for a tube not being drawn according to the protocol, and any comments about the venipuncture.

Note from the Obesity/Adiposity WG: The investigator should record whether the blood was drawn and whether the full amount was obtained.

Process the Sample for the Serum

Editor’s Note: Please review chapter 8 of the Laboratory Procedures Manual from the NHANES 2007-2008 for a full description of blood processing procedures: 2007-2008 NHANES Lab Manual.

  • Allow the blood to clot by setting aside for 30-45 minutes at room temperature. Do not clot for more than 1 hour.
  • Centrifuge the tube at room temperature to separate the serum and aliquot into an appropriate storage tube.
  • Determine if the serum is hemolyzed, turbid, lipemic, or icteric. If so, enter a comment to describe the plasma

Laboratory Assay for Adipocytokines

There are now many laboratory assays that have been approved by the U.S. Food and Drug Administration (FDA). Once an assay is chosen for a particular study, the WG recommends that consistent methods are followed so that assay results will be comparable.

Personnel and Training Required

Certified phlebotomist

Laboratory capable of performing required assays

Equipment Needs

Phlebotomy supplies

Requirements
Requirement CategoryRequired
Major equipment No
Specialized training No
Specialized requirements for biospecimen collection No
Average time of greater than 15 minutes in an unaffected individual No
Mode of Administration

Bioassay

Lifestage

Child, Adolescent, Adult

Participants

All ages: Child-adult

Selection Rationale

Adipocytokines reflect consequential pathways of adiposity. The ELISA method has high feasibility, relatively low cost, and low subject burden.

Language

English

Standards
StandardNameIDSource
Human Phenotype Ontology Obesity HP:0001513 HPO
caDSR Form PhenX PX230201 - Adipocytokines 6872850 caDSR Form
Derived Variables

None

Process and Review

Not applicable.

Protocol Name from Source

National Health and Nutrition Examination Survey Questionnaire (NHANES), Laboratory Procedures Manual, 2007

Source

Centers for Disease Control and Prevention (CDC). National Center for Health Statistics (NCHS) (2007). National Health and Nutrition Examination Survey Questionnaire. Laboratory Procedures Manual. Hyattsville, MD: U.S. Department of Health and Human Services, Centers for Disease Control and Prevention.

General References

www.emdmillipore.com/US/en/product/Human-Adiponectin-ELISA,MM_NF-EZHADP-61K

www.emdmillipore.com/US/en/product/Human-Leptin-%22Dual-Range%22-ELISA,MM_NF-EZHL-80SK

Protocol ID

230201

Variables
Export Variables
Variable Name Variable IDVariable DescriptiondbGaP Mapping
PX230201_Adipocytokines_Assay_Repeatability
PX230201080000 Repeatability of the assay N/A
PX230201_Adipocytokines_Assay_Results
PX230201100000 Adipocytokines assay results N/A
PX230201_Adipocytokines_Assay_Type
PX230201060000 Record the type of assay used for more
adipocytokines testing. show less
N/A
PX230201_Adipocytokines_Blood_Draw_Comments
PX230201040200 Record any comments about the blood draw, more
including any reasons for the tube not being drawn according to the protocol. show less
N/A
PX230201_Adipocytokines_Blood_Draw_Done
PX230201040000 Was blood drawn? N/A
PX230201_Adipocytokines_Blood_Draw_Sample
PX230201040100 Was full amount obtained? N/A
PX230201_Adipocytokines_Chemotherapy
PX230201020000 Have you received cancer chemotherapy in the more
past four weeks or do you anticipate such therapy in the next four weeks? (exclusion from Phlebotomy) show less
N/A
PX230201_Adipocytokines_Coefficient_Of_Variation
PX230201090000 Coefficient of variation for the assay N/A
PX230201_Adipocytokines_Equipment_Make
PX230201070100 Make of the equipment used to perform the more
adipocytokines assay. show less
N/A
PX230201_Adipocytokines_Equipment_Manufacturer
PX230201070200 Manufacturer of the equipment used to more
perform the adipocytokines assay. show less
N/A
PX230201_Adipocytokines_Exclusion_Criteria
PX230201030000 Exclusion Criteria N/A
PX230201_Adipocytokines_Hemophilia
PX230201010000 Do you have hemophilia? (exclusion from more
Phlebotomy) show less
N/A
PX230201_Adipocytokines_Sample_Comments
PX230201050000 Record any comments about the sample during more
processing. show less
N/A
Obesity
Measure Name

Adipocytokines

Release Date

September 9, 2015

Definition

These are bioassays to measure high molecular weight adiponectin.

Purpose

Leptin human ELISA kits are designed to quantify protein levels of human leptin in serum, plasma, supernatant, and other biological fluids. Leptin is a hormone secreted by adipocytes.

Adiponectin is another adipocyte-specific protein which plays a role in the development of insulin resistance and atherosclerosis. HMW adiponectin is the primary bioactive form with insulin-sensitizing activity and is recommended for measurement. Its circulating levels negatively correlate with obesity, insulin resistance, and coronary artery disease.

Keywords

Adipocytokines, leptin, adiponectin, high molecular weight adiponectin, HMWA, obesity, insulin resistance, National Health and Nutrition Examination Survey, NHANES

Measure Protocols
Protocol ID Protocol Name
230201 Adipocytokines
Publications

There are no publications listed for this protocol.