Protocol - Haptoglobin Level

Description

This protocol provides instructions for drawing, processing, and storing blood according to the National Health and Nutrition Examination Survey (NHANES) methods. Because there are many comparable assays for measuring haptoglobin, the protocol also provides basic guidelines to aid comparability among different studies.

Specific Instructions

The National Health and Nutrition Examination Survey (NHANES) instructions for drawing, processing and storing blood provide a standard methodology used successfully for many years to ensure comparable results across study sites. However, the Sickle Cell Disease Cardiovascular, Pulmonary, and Renal Working Group notes that certain aspects (e.g., exclusion criteria) of the NHANES protocol are study specific and might not be applicable to all types of studies (e.g., sickle cell disease). Investigators who want to include participants that have hemophilia or have received cancer chemotherapy in the last 4 weeks will need to implement special venipuncture procedures.

In general, haptoglobin levels can be combined with other indirect markers of hemolysis (Aspartate Aminotransferase Level, Reticulocyte Count, Lactate Dehydrogenase Level, and Bilirubin Level) to derive a hemolytic component. However, in sickle cell disease patients, the haptoglobin level is likely to be undetectable. Accordingly, once a patient has been diagnosed with sickle cell disease, the Working Group does not recommend repeating haptoglobin measurements.

Haptoglobin levels are a useful indicator of hemolysis because serum concentrations decrease when hemoglobin is released from red cells. However, the Working Group notes that haptoglobin levels can be altered by other factors. For example, haptoglobin levels will be increased with acute inflammation and decreased with liver dysfunction. Haptoglobin levels can also be reduced due to genetic variation. A number of medications may also increase or decrease haptoglobins

Determining haptoglobin levels via a serum sample is preferred. However, the Working Group acknowledges that collection of blood samples for the measurement of analytes requires a general determination of whether to use serum or plasma for the assay and also a determination of the type of collection tube to be obtained. For example, if serum is to be used, a determination needs to be made as to whether red top or serum gel separator collection tubes are used. While comparable values are obtained for many analytes from either serum or plasma, there may be situations where differences are more pronounced and serum- or plasma-specific norms will be needed for references. The NHANES protocol presented here uses red top/serum separator tubes. At times it may be possible to collect both, but other considerations such as participant burden may be the deciding factor. It is important to match assay type with sample type. Some automated devices may preclude the use of serum, for example, while others may be optimized for it. Investigators should choose methods of collection that match the methods of analysis. This will best be done by communicating with the laboratory where the proposed assays will be performed. They will become an important partner with you in ensuring that there is compatibility from collection to assays to interpretation and reporting of levels and results.

The Sickle Cell Disease Curative Therapy Working Group recommends that haptoglobin levels be determined in sickle cell patients undergoing hematopoietic cell transplant at one time point pre-transplant (baseline) and 100 days, six months, and annually post- transplant.

Availability

Available

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Protocol

The following is a summary version of the full National Health and Nutrition Examination Survey 2011-2012 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 SP answers "No" or "Don’t Know," blood is drawn from the SP.

2. Have you received cancer chemotherapy in the past four weeks or do you anticipate such therapy in the next four 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 SP answers "No" or "Don’t Know," blood is drawn from the SP.

Venipuncture Procedures

Editor’s Note: Please review chapter 4 of the Laboratory Procedures Manual from the 2011-2012 National Health and Nutrition Examination Survey (NHANES) for a full description of phlebotomy procedures. This manual is posted here, and is also available at the NHANES website: 2011-2012 NHANES Laboratory Procedures Manual.

Venipuncture should generally be performed using the median cubital, cephalic, or basilic 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 the 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.

Blood Processing

Please review chapter 8 of the Laboratory Procedures Manual from the National Health and Nutrition Examination Survey 2011-2012 for a full description of blood processing procedures: 2011-2012 NHANES Laboratory Procedures Manual.

Allow the blood to clot by setting aside for 30 to 45 minutes at room temperature. Do not clot for more than an 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 serum.

Laboratory Assay for Haptoglobin

The Sickle Cell Disease Cardiovascular, Pulmonary, and Renal Working Group notes that there are a number of different assays and instruments that are appropriate to measure the concentration of haptoglobin. Once an assay is chosen for a particular study, the Working Group recommends that no changes in the protocol be made over the course of the study. To aid comparability, the Working Group recommends that the investigator record the make and manufacturer of equipment used and the repeatability and coefficients of variation for the assay.

Reference Ranges for Haptoglobin:

In normal individuals, haptoglobin levels range between 45 and 165 milligrams per deciliter of blood (mg/dL). Levels lower than 45 mg/dL can indicate increased rate of red blood cell death.

Personnel and Training Required

Phlebotomist

Equipment Needs

Laboratory with the ability to perform the haptoglobin assay.

Requirements

Requirement Category	Required
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

Toddler, Child, Adolescent, Adult, Senior, Pregnancy

Participants

Individuals age 1 year and older.

Selection Rationale

The Sickle Cell Disease Cardiovascular, Pulmonary, and Renal Working Group selected the National Health and Nutrition Examination Survey 2011-2012 protocol as the best standardized methodology for blood collection, processing, and storage.

Language

English

Standards

Standard	Name	ID	Source
Human Phenotype Ontology	Sickle Cell Anemia	ORPHA:232	HPO
Human Phenotype Ontology	Anemia	OMIM:603903	HPO
caDSR Form	PhenX PX810801 - Haptoglobin Level	6252679	caDSR Form

Derived Variables

Hemolytic Component:

Haptoglobin levels can be combined with other indirect markers of hemolysis (Bilirubin Level, Reticulocyte Count, Aspartate Aminotransferase Level, and Lactate Dehydrogenase Level) by principle component analysis (PCA) to derive a hemolytic component for sickle cell disease patients.

Nouraie, M., Lee, J. S., Zhang, Y., Kanias, T., Zhao, X., Xiong, Z., Oriss, T. B., Zeng, Q., Kato, G. J., Gibbs, J. S., Hildesheim, M. E., Sachdev, V., Barst, R. J., Machado, R. F., Hassell, K. L., Little, J. A., Schraufnagel, D. E., Krishnamurti, L., Novelli, E., Girgis, R. E., Morris, C.R., Rosenzweig, E. B., Badesch, D. B., Lanzkron, S., Castro, O. L., Goldsmith, J. C., Gordeuk, V. R., Galdwin, M. T., & Walk-PHASST Investigators and Patients. (2013). The relationship between the severity of hemolysis, clinical manifestations and risk of death in 415 patients with sickle cell anemia in the US and Europe. Haematologica, 98(3), 464-472.

Process and Review

Not applicable.

Protocol Name from Source

National Health and Nutrition Examination Survey Questionnaire (NHANES), Laboratory Procedures Manual, 2011

Source

Centers for Disease Control and Prevention (CDC). (2011). 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.

Children’s Hospital Los Angeles, Laboratory Guide - Lexicomp Online, 2015

General References

MedlinePlus: www.nlm.nih.gov/medlineplus/ency/article/003634.htm

Potoka, K. P., & Gladwin, M. T. (2015). Vasculopathy and pulmonary hypertension in sickle cell disease. American Journal of Physiology - Lung Cellular and Molecular Physiology, 308, L314-L324.

Reiter, C.D., Wang, X., Tanus-Santos, J.E., Hogg, N., Cannon, R.O. 3rd, Schechter, A.N., & Gladwin, M.T. (2002). Cell-free hemoglobin limits nitric oxide bioavailability in sickle-cell disease. Nature Medicine, 8(12), 1383-1389.

Sachdev, V., Kato, G. J., Gibbs, J. S., Barst, R. J., Machado, R. F., Nouraie, M., Hassell, K. L., Little, J. A., Schraufnagel, D. E., Krishnamurti, L., Novelli, E. M., Girgis, R. E., Morris, C. R., Rosenzweig, E. B., Badesch, D. B., Lanzkron, S., Castro, O. L., Taylor, J. G. 6th, Hannoush, H., Goldsmith, J. C., Gladwin, M. T., Gordeuk, V. R., & Walk-PHASST Investigators. (2011). Echocardiographic markers of elevated pulmonary pressure and left ventricular diastolic dysfunction are associated with exercise intolerance in adults and adolescents with homozygous sickle cell anemia in the United States and United Kingdom. Circulation, 124(13), 1452-1460.

Protocol ID

810801

Variables

Export Variables

Variable Name	Variable ID	Variable Description	dbGaP Mapping
PX810801_Haptoglobin_Amount
	PX810801080000	Haptoglobin level	N/A
PX810801_Haptoglobin_Assay_Repeatability
	PX810801110000	Repeatability of the haptoglobin assay.	N/A
PX810801_Haptoglobin_Blood_DrawTube_Deviation
	PX810801040000	Record reasons for a tube not being drawn more according to the protocol. show less	N/A
PX810801_Haptoglobin_Blood_Draw_Results
	PX810801030000	Record the results of the blood draw.	N/A
PX810801_Haptoglobin_Chemotheraphy_4weeks
	PX810801020000	Have you received cancer chemotherapy in the more past four weeks or do you anticipate such therapy in the next four weeks? show less	Variable Mapping
PX810801_Haptoglobin_Coefficients_Variation_Assay
	PX810801120000	Coefficients of variation of the haptoglobin more assay. show less	N/A
PX810801_Haptoglobin_Equipment_Make
	PX810801090000	Make of the equipment used to perform the more haptoglobin assay. show less	N/A
PX810801_Haptoglobin_Equipment_Manufacturer
	PX810801100000	Manufacturer of the equipment used to more perform the haptoglobin assay. show less	N/A
PX810801_Haptoglobin_Hemophilia
	PX810801010000	Do you have hemophilia?	Variable Mapping
PX810801_Haptoglobin_Serum_Description
	PX810801070000	If serum is hemolyzed, turbid, lipemic, or more iteric, then describe. show less	N/A
PX810801_Haptoglobin_Serum_Determination
	PX810801060000	Determine if the serum is hemolyzed, turbid, more lipemic, or icteric. show less	N/A
PX810801_Haptoglobin_Venipuncture_Comments
	PX810801050000	Record any comments about the venipuncture.	N/A

SCD Cardiovascular, Pulmonary, and Renal

Measure Name

Haptoglobin Level

Release Date

July 30, 2015

Definition

A bioassay to measure haptoglobin, which is produced by the liver and transports hemoglobin to the liver for recycling.

Purpose

Low levels of haptoglobin are an indirect marker of hemolysis.

Keywords

Haptoglobin, hemolysis, hemoglobin, sickle cell disease, SCD, anemia, hypoxemia, hemolytic anemia, Liver, red blood cells, pulmonary hypertension, pH, chronic kidney disease, CKD, vasculopathy, stroke, hemolytic component, "Cardiovascular, Pulmonary, and Renal"

Measure Protocols

Protocol ID	Protocol Name
810801	Haptoglobin Level

Publications

There are no publications listed for this protocol.

Mapping for PX810801_Haptoglobin_Chemotheraphy_4weeks

dbGAP Study	dbGAP Variable	dbGAP Variable Description
Genotype-Tissue Expression (GTEx)	phv00169161	Hemophilia (Potential Exposure: Sexual Activity)

Mapping for PX810801_Haptoglobin_Hemophilia

dbGAP Study	dbGAP Variable	dbGAP Variable Description
Genome Wide Association Study of Chronic TMD: Discovery Phase	phv00260451	MED 2. Hematologic: hemophilia/other bleeding disorders.
Genotype-Tissue Expression (GTEx)	phv00169160	Hemophilia (Information)
Genotype-Tissue Expression (GTEx)	phv00169161	Hemophilia (Potential Exposure: Sexual Activity)