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Protocol - Reticulocyte Count

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Protocol Name from Source:

This section will be completed when reviewed by an Expert Review Panel.

Availability:

Publicly available

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 and instruments for measuring reticulocyte count, the protocol also provides basic guidelines to aid comparability among different studies.

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 [link[www.phenx.org/Portals/0/phenx-content/Supplements/SickleCell/WG1/DocumentsForReview/Documents_for_Additional_Information/2011-12_Laboratory_Procedures_Manual.pdf|here]], and is also available at the NHANES website: [link[www.cdc.gov/nchs/data/nhanes/nhanes_11_12/2011-12_Laboratory_Procedures_Manual.pdf|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: [link[www.cdc.gov/nchs/data/nhanes/nhanes_11_12/2011-12_Laboratory_Procedures_Manual.pdf|2011-2012 NHANES Laboratory Procedures Manual]]

Fill a 3 or 4 ml K3 EDTA tube with blood.

Laboratory Assay for Reticulocyte Count

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 reticulocyte count from serum. 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 Reticulocyte Count:

Children: 3 - 6%

Adults: 0.5 - 1.5%

Percentage of reticulocytes and absolute reticulocyte counts (ARC) are given by the following formulas:

Reticulocyte Percentage = [Number of Reticulocytes (thousands/µL) x RBC count (millions/µL)] X 100

Absolute reticulocyte count (thousands/µL) = reticulocyte % x RBC count (millions/µL) x 10

Personnel and Training Required

Phlebotomist

Equipment Needs

Laboratory with the ability to perform the reticulocyte count assay.

Requirements

Requirement CategoryRequired
Average time of greater than 15 minutes in an unaffected individualNo
Major equipmentNo
Specialized requirements for biospecimen collectionNo
Specialized trainingNo

Mode of Administration

Self-administered

Life Stage:

Toddler, Child, Adolescent, Adult, Senior, Pregnancy

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.

Reticulocyte count can be combined with other indirect markers of hemolysis ([mlink[811200|Aspartate Aminotransferase Level]], [mlink[810800|Haptoglobin Level]], [mlink[811000|Lactate Dehydrogenase Level]], and [mlink[810900|Bilirubin Level]]) to derive a hemolytic component for sickle cell disease patients.

Reticulocyte count analysis is performed on anticoagulated blood, collected in an EDTA tube.

Research Domain Information

Release Date:

July 30, 2015

Definition

A bioassay to measure the number of reticulocytes (immature red blood cells).

Purpose

Reticulocyte count measures how fast red blood cells are made. Elevated reticulocyte counts are associated with blood loss, hemolysis and hemolytic anemia, and in response to the anemia of clinical conditions such as sickle cell disease. Reticulocyte counts can also be used to measure efficacy of treatments (e.g., iron replacement therapy).

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

StandardNameIDSource
Common Data Elements (CDE)Hematology Reticulocyte Laboratory Total Value2632396CDE Browser

Process and Review

This section will be completed when reviewed by an Expert Review Panel.

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.

General References

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. doi: 10.3324/haematol.2012.068965

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

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:

810601

Variables:

Export Variables
Variable NameVariable IDVariable DescriptionVersiondbGaP Mapping
PX810601_Reticulocyte_HemophiliaPX810601010000Do you have hemophilia?4N/A
PX810601_Reticulocyte_Chemotheraphy_4weeksPX810601020000Have you received cancer chemotherapy in the past four weeks or do you anticipate such therapy in the next four weeks?4N/A
PX810601_Reticulocyte_AgePX810601030000Age4N/A
PX810601_Reticulocyte_Blood_Draw_ResultsPX810601040000Record the results of the blood draw.4N/A
PX810601_Reticulocyte_Blood_Draw_Tube_DeviationPX810601050000Record reasons for a tube not being drawn according to the protocol.4N/A
PX810601_Reticulocyte_Venipuncture_CommentsPX810601060000Record any comments about the venipuncture.4N/A
PX810601_Reticulocyte_AmountPX810601070000Number of reticulocytes4N/A
PX810601_Reticulocyte_Equipment_MakePX810601080000Make of the equipment used to perform the reticulocyte count assay.4N/A
PX810601_Reticulocyte_Equipment_ManufacturerPX810601090000Manufacturer of the equipment used to perform the reticulocyte count assay.4N/A
PX810601_Reticulocyte_Assay_RepeatabilityPX810601100000Repeatability of the reticulocyte count assay.4N/A
PX810601_Reticulocyte_Coefficients_Variation_AssayPX810601110000Coefficients of variation of the reticulocyte count assay.4N/A