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Department of Medicine, William Beaumont Army Medical Center, El Paso, TX 79920.
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Department of Medicine, Washington Hospital Center, and
Medlantic Research Institute, Washington, DC 20302.
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Department of Medicine, Johns Hopkins University,
Baltimore, MD 21215.
Departments of
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Medicine,
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Pediatrics, and
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Clinical Investigation, Walter Reed Army Medical Center,
Washington, DC 20307.
a Author for correspondence. Fax 202-782-3881; e-mail rmtuttle{at}hotmail.com
Background: Thyroglobulin mRNA can be detected qualitatively in the peripheral blood of patients with metastatic thyroid cancer, thyroid cancer patients with residual thyroid bed uptake, and individuals with no known thyroid disease with intact thyroid glands by use of a lengthy, highly sensitive extraction technique. To improve and broaden the clinical usefulness of this assay, we developed a quantitative reverse transcription (RT)-PCR assay for thyroglobulin mRNA, using RNA recovered from whole blood with a simplified extraction technique.
Methods: Whole blood was drawn from 32 healthy subjects in standard EDTA blood collection tubes. Total RNA was extracted from whole blood, using the PUREscript RNA Isolation Kit. RT-PCR using intron-spanning primers was used to quantitatively amplify thyroglobulin mRNA, using the ABI PRISM 7700 Sequence Detection System with a fluorescent-labeled, thyroglobulin-specific oligonucleotide probe. Thyroid RNA calibration curves were created using total RNA recovered from a single nondiseased thyroid gland.
Results: Qualitative RT-PCR demonstrated the presence of thyroglobulin mRNA in the whole blood sample of each healthy subject. The mean concentration of thyroglobulin mRNA detected in these subjects was 433 ± 69 ng of total thyroid RNA per liter of whole blood (range, 26–1502 ng/L). Overall assay imprecision (CV) was 24% for five samples analyzed 10 times each in separate analytical runs on different days.
Conclusions: Thyroglobulin mRNA can be accurately detected and quantified in peripheral blood from healthy subjects. This new quantitative technique may improve the clinical utility of circulating thyroglobulin mRNA detection in patients with thyroid disease.© 1999 American Association for Clinical Chemistry
The following articles in journals at HighWire Press have cited this article:
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 Q. He, J.-P. Wang, M. Osato, and L. B. Lachman Real-Time Quantitative PCR for Detection of Helicobacter pylori J. Clin. Microbiol., October 1, 2002; 40(10): 3720 - 3728. [Abstract] [Full Text] [PDF]
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M. K. Gupta, L. Taguba, R. Arciaga, A. Siperstein, C. Faiman, A. Mehta, and S. S. K. Reddy Detection of Circulating Thyroid Cancer Cells by Reverse Transcription-PCR for Thyroid-stimulating Hormone Receptor and Thyroglobulin: The Importance of Primer Selection Clin. Chem., October 1, 2002; 48(10): 1862 - 1865. [Full Text] [PDF]
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L. Fugazzola, A. Mihalich, L. Persani, N. Cerutti, M. Reina, M. Bonomi, E. Ponti, D. Mannavola, E. Giammona, G. Vannucchi, et al. Highly Sensitive Serum Thyroglobulin and Circulating Thyroglobulin mRNA Evaluations in the Management of Patients with Differentiated Thyroid Cancer in Apparent Remission J. Clin. Endocrinol. Metab., July 1, 2002; 87(7): 3201 - 3208. [Abstract] [Full Text] [PDF]
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F. Savagner, P. Rodien, P. Reynier, V. Rohmer, J.-C. Bigorgne, and Y. Malthiery Analysis of Tg Transcripts by Real-Time RT-PCR in the Blood of Thyroid Cancer Patients J. Clin. Endocrinol. Metab., February 1, 2002; 87(2): 635 - 639. [Abstract] [Full Text] [PDF]
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