A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate

Maria Rossing; Bogumil Kaczkowski; Ewa Futoma-Kazmierczak; Martin Glud; Mikkel Klausen; Jens Faber; Birte Nygaard; Katalin Kiss; Christian Hjort Sørensen; Finn Cilius Nielsen; Finn Noe Bennedbæk; Lennart Jan Friis-Hansen

doi:10.3109/00365513.2010.522250

A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate

Maria Rossing, Bogumil Kaczkowski, Ewa Futoma-Kazmierczak, Martin Glud, Mikkel Klausen, Jens Faber, Birte Nygaard, Katalin Kiss, Christian Hjort Sørensen, Finn Cilius Nielsen, Finn Noe Bennedbæk, Lennart Jan Friis-Hansen

6 Citations (Scopus)

Abstract

Context. microRNA (miRNA) expression profiling and classification of tissue obtained from fine-needle aspirates (FNA) could be a major improvement of the preoperative diagnosis of thyroid nodules. Objective. Before this can be clinically implemented, a robust and non-toxic method for obtaining sufficient quantity and quality of RNA from single in vivo FNA has to be established. RNAlater is a non-toxic RNA-stabilizing agent. However, due to the high density of RNAlater, pelleting of the tissue samples is difficult, and results in low recovery of RNA that is insufficient for subsequent miRNA array expression analysis. We therefore developed a simple centrifugation method for capturing tissue stored in RNAlater on a 0.45-μm filter. Design. FNA from 24 patients with a solitary cold thyroid nodule was stored in Trizol, liquid nitrogen, or RNAlater. The tissue stored in RNAlater was either pelleted by centrifugation or captured on the 0.45-μm filters. RNA was extracted using the Trizol method. To validate results, FNA from additional 30 patients were analyzed based on the modified RNAlater protocol. Main outcome. Capturing FNA tissue samples on the filters increased the RNA yield 10 fold and maintained RNA purity, permitting miRNA array expression profiling and allowing comparable levels of known miRNA-clusters regardless of preservation technique. Results were confirmed in an additional 30 patients. Conclusion. The modified RNAlater protocol is well suited for isolating RNA from single thyroid in vivo FNA in a clinical setting. Furthermore this permits shipping of FNA samples at room temperature from peripheral centers to a centralized array core facility.

Original language	English
Journal	Scandinavian Journal of Clinical & Laboratory Investigation
Volume	70
Issue number	8
Pages (from-to)	529-534
Number of pages	6
ISSN	0036-5513
DOIs	https://doi.org/10.3109/00365513.2010.522250
Publication status	Published - Dec 2010

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Rossing, M., Kaczkowski, B., Futoma-Kazmierczak, E., Glud, M., Klausen, M., Faber, J., Nygaard, B., Kiss, K., Sørensen, C. H., Nielsen, F. C., Bennedbæk, F. N., & Friis-Hansen, L. J. (2010). A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate. Scandinavian Journal of Clinical & Laboratory Investigation, 70(8), 529-534. https://doi.org/10.3109/00365513.2010.522250

A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate. / Rossing, Maria; Kaczkowski, Bogumil; Futoma-Kazmierczak, Ewa et al.
In: Scandinavian Journal of Clinical & Laboratory Investigation, Vol. 70, No. 8, 12.2010, p. 529-534.

Research output: Contribution to journal › Journal article › Research › peer-review

Rossing, M, Kaczkowski, B, Futoma-Kazmierczak, E, Glud, M, Klausen, M, Faber, J, Nygaard, B, Kiss, K, Sørensen, CH, Nielsen, FC, Bennedbæk, FN & Friis-Hansen, LJ 2010, 'A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate', Scandinavian Journal of Clinical & Laboratory Investigation, vol. 70, no. 8, pp. 529-534. https://doi.org/10.3109/00365513.2010.522250

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title = "A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate",

abstract = "Context. microRNA (miRNA) expression profiling and classification of tissue obtained from fine-needle aspirates (FNA) could be a major improvement of the preoperative diagnosis of thyroid nodules. Objective. Before this can be clinically implemented, a robust and non-toxic method for obtaining sufficient quantity and quality of RNA from single in vivo FNA has to be established. RNAlater is a non-toxic RNA-stabilizing agent. However, due to the high density of RNAlater, pelleting of the tissue samples is difficult, and results in low recovery of RNA that is insufficient for subsequent miRNA array expression analysis. We therefore developed a simple centrifugation method for capturing tissue stored in RNAlater on a 0.45-μm filter. Design. FNA from 24 patients with a solitary cold thyroid nodule was stored in Trizol, liquid nitrogen, or RNAlater. The tissue stored in RNAlater was either pelleted by centrifugation or captured on the 0.45-μm filters. RNA was extracted using the Trizol method. To validate results, FNA from additional 30 patients were analyzed based on the modified RNAlater protocol. Main outcome. Capturing FNA tissue samples on the filters increased the RNA yield 10 fold and maintained RNA purity, permitting miRNA array expression profiling and allowing comparable levels of known miRNA-clusters regardless of preservation technique. Results were confirmed in an additional 30 patients. Conclusion. The modified RNAlater protocol is well suited for isolating RNA from single thyroid in vivo FNA in a clinical setting. Furthermore this permits shipping of FNA samples at room temperature from peripheral centers to a centralized array core facility.",

author = "Maria Rossing and Bogumil Kaczkowski and Ewa Futoma-Kazmierczak and Martin Glud and Mikkel Klausen and Jens Faber and Birte Nygaard and Katalin Kiss and S{\o}rensen, {Christian Hjort} and Nielsen, {Finn Cilius} and Bennedb{\ae}k, {Finn Noe} and Friis-Hansen, {Lennart Jan}",

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T1 - A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate

AU - Rossing, Maria

AU - Kaczkowski, Bogumil

AU - Futoma-Kazmierczak, Ewa

AU - Glud, Martin

AU - Klausen, Mikkel

AU - Faber, Jens

AU - Nygaard, Birte

AU - Kiss, Katalin

AU - Sørensen, Christian Hjort

AU - Nielsen, Finn Cilius

AU - Bennedbæk, Finn Noe

AU - Friis-Hansen, Lennart Jan

PY - 2010/12

Y1 - 2010/12

N2 - Context. microRNA (miRNA) expression profiling and classification of tissue obtained from fine-needle aspirates (FNA) could be a major improvement of the preoperative diagnosis of thyroid nodules. Objective. Before this can be clinically implemented, a robust and non-toxic method for obtaining sufficient quantity and quality of RNA from single in vivo FNA has to be established. RNAlater is a non-toxic RNA-stabilizing agent. However, due to the high density of RNAlater, pelleting of the tissue samples is difficult, and results in low recovery of RNA that is insufficient for subsequent miRNA array expression analysis. We therefore developed a simple centrifugation method for capturing tissue stored in RNAlater on a 0.45-μm filter. Design. FNA from 24 patients with a solitary cold thyroid nodule was stored in Trizol, liquid nitrogen, or RNAlater. The tissue stored in RNAlater was either pelleted by centrifugation or captured on the 0.45-μm filters. RNA was extracted using the Trizol method. To validate results, FNA from additional 30 patients were analyzed based on the modified RNAlater protocol. Main outcome. Capturing FNA tissue samples on the filters increased the RNA yield 10 fold and maintained RNA purity, permitting miRNA array expression profiling and allowing comparable levels of known miRNA-clusters regardless of preservation technique. Results were confirmed in an additional 30 patients. Conclusion. The modified RNAlater protocol is well suited for isolating RNA from single thyroid in vivo FNA in a clinical setting. Furthermore this permits shipping of FNA samples at room temperature from peripheral centers to a centralized array core facility.

AB - Context. microRNA (miRNA) expression profiling and classification of tissue obtained from fine-needle aspirates (FNA) could be a major improvement of the preoperative diagnosis of thyroid nodules. Objective. Before this can be clinically implemented, a robust and non-toxic method for obtaining sufficient quantity and quality of RNA from single in vivo FNA has to be established. RNAlater is a non-toxic RNA-stabilizing agent. However, due to the high density of RNAlater, pelleting of the tissue samples is difficult, and results in low recovery of RNA that is insufficient for subsequent miRNA array expression analysis. We therefore developed a simple centrifugation method for capturing tissue stored in RNAlater on a 0.45-μm filter. Design. FNA from 24 patients with a solitary cold thyroid nodule was stored in Trizol, liquid nitrogen, or RNAlater. The tissue stored in RNAlater was either pelleted by centrifugation or captured on the 0.45-μm filters. RNA was extracted using the Trizol method. To validate results, FNA from additional 30 patients were analyzed based on the modified RNAlater protocol. Main outcome. Capturing FNA tissue samples on the filters increased the RNA yield 10 fold and maintained RNA purity, permitting miRNA array expression profiling and allowing comparable levels of known miRNA-clusters regardless of preservation technique. Results were confirmed in an additional 30 patients. Conclusion. The modified RNAlater protocol is well suited for isolating RNA from single thyroid in vivo FNA in a clinical setting. Furthermore this permits shipping of FNA samples at room temperature from peripheral centers to a centralized array core facility.

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DO - 10.3109/00365513.2010.522250

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SN - 0036-5513

VL - 70

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EP - 534

JO - Scandinavian Journal of Clinical & Laboratory Investigation

JF - Scandinavian Journal of Clinical & Laboratory Investigation

IS - 8

ER -

A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate

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