Protocol - Thyroid Stimulating Hormone (hTSH) in Serum

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This protocol provides an overview of the thyroid-stimulating hormone immunoassay according to the local laboratory. Some examples include LabCorp test 004259 or Quest Diagnostics test 899. Often free thyroxine (free T4 or FT4) level is tested along with TSH (LabCorp test 224576 or Quest Diagnostics test 867; CPT 84439). Because there are many comparable assays and instruments for measuring TSH, the protocol also provides basic guidelines to aid comparability among different studies.

Specific Instructions

Participants should stop taking biotin (vitamin B7, vitamin B8, vitamin H, coenzyme R) at least 72 hours prior to sample collection.

The Sickle Cell Disease Curative Therapy Working Group also recommends collecting free T4 (Thyroxine). When used concurrently with Thyroid-stimulating Hormone, free T4 provides more information about the etiology of hypothyroidism.




Specimen Requirements and Storage Instructions

The thyroid-stimulating hormone assay is performed on serum. 0.3 to 0.8 milliliters of serum should be collected in a red-top or gel-barrier tube. The sample is stable for 14 days at room temperature, refrigerated or frozen. 

Summary of the Thyroid-Stimulating Hormone Assay

The sample is incubated with biotinylated and ruthenium labeled monoclonal TSH-specific antibodies to form a sandwich complex. Voltage is applied to the sample to induce a chemiluminescence which is measured by a photomultiplier. Results are measured against a calibration curve.

Thyroid-Stimulating Hormone Assay Reference Ranges


Range (micro-International Units per milliliter – ulU/mL)

0 to 6 d


7 d to 3 m


3 m 1 d to 12 m


1 to 5 y


6 to 10 y


>10 y


The Sickle Cell Disease Curative Therapies Working Group notes that there are a number of different assays and instruments that are appropriate to measure thyroid stimulating hormone. 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 name of the assay, the make and manufacturer of equipment used and the repeatability and coefficients of variation for the assay.

Personnel and Training Required

A phlebotomist is required to draw the blood sample.

Equipment Needs

Laboratory with the equipment needed to perform the thyroid stimulating hormone assay

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



Infant, Toddler, Child, Adolescent, Adult


All ages

Selection Rationale

The TSH test is widely available, standardized methodology for collecting, processing, and storing blood, performing the prothrombin test, and reporting results.



Derived Variables


Process and Review

Not applicable.

Protocol Name from Source

Thyroid Stimulating Hormone (TSH), CPT 84443


Labcorp. (n.d.). Thyroid-stimulating Hormone (TSH). Retrieved February 23, 2021, from https://www.labcorp.com/tests/004259/thyroid-stimulating-hormone-tsh

Quest Diagnostics. TSH. Retrieved July 16, 2021, from TSH | Test Detail | Quest Diagnostics

General References

ElAlfy, M. S., El-Sherif, N. H., Sakr, H. M., & El Ashkar, M. (2019). Thyroid hemodynamic alterations in Egyptian patients with sickle cell disease: Relation to disease severity, total body iron and thyroid function. Expert Review of Hematology, 12(8), 589–596.

Farmakis, D., Angastiniotis, M., & Eleftheriou, A. (2017). A short guide for the management of transfusion dependent thalassaemia (TDT) (TIF Publication No. 23). Thalassaemia International Federation.

Hagag, A. A., El-Asy, H. M., Badraia, I. M., Hablas, N. M., & El-Latif, A. (2019). Thyroid function in Egyptian children with sickle cell anemia in correlation with iron load. Endocrine, Metabolic & Immune Disorders Drug Targets, 19(1), 46–52.

Protocol ID


Export Variables
Variable Name Variable IDVariable DescriptiondbGaP Mapping
PX851101010000 Was 0.3 to 0.8 milliliters of serum more
collected in a red-top or gel-barrier tube? show less
PX851101050000 Make and manufacturer of equipment N/A
PX851101060000 What is the repeatability and coefficients more
of variation of the assay? show less
PX851101040000 Were the results measured against a more
calibration curve? show less
PX851101030000 Was the sample incubated with biotinylated more
and ruthenium labeled monoclonal TSH-specific antibodies to form a sandwich complex? show less
PX851101020000 Was the sample is stable for 14 days at room more
temperature, refrigerated or frozen? show less
Curative Therapies
Measure Name

Thyroid Stimulating Hormone

Release Date

August 16, 2021


Thyroid stimulating hormone is produced by the pituitary gland and stimulates release of thyroxine from the thyroid.


This measure is used to assess thyroid status, including screening for hyperthyroidism and hypothyroidism. Complications of iron overload in sickle cell disease contribute to iron-related morbidity and mortality. Chronic red blood cell transfusions in sickle cell disease can lead to iron accumulation in the endocrine system and disrupt the secretion of thyroid stimulating hormone. 


sickle cell, sickle cell disease, thyroid stimulating hormone, TSH, iron, iron overload, curative therapies, blood transfusion, free T4, endocrine system, National Health and Nutrition Examination Survey, NHANES

Measure Protocols
Protocol ID Protocol Name
851101 Thyroid Stimulating Hormone (hTSH) in Serum

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