Comparative stabilities of quantitative human immunodeficiency virus RNA in plasma from samples collected in VACUTAINER CPT, VACUTAINER PPT, and standard VACUTAINER tubes.

Author(s): Holodniy M, Mole L, Yen-Lieberman B, Margolis D, Starkey C, Carroll R, Spahlinger T, Todd J, Jackson JB

Publication: J Clin Microbiol, 1995, Vol. 33, Page 1562-6

PubMed ID: 7650187 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of collection tube type, anticoagulant, delayed centrifugation, and storage time and temperature on human immunodeficiency virus (HIV) RNA levels in plasma and serum.

Conclusion of Paper

The highest levels of HIV RNA were recovered when specimens were collected in either Vacutainers or cell preparation tubes (CPT) containing potassium EDTA. The lowest levels of HIV RNA were measured in specimens collected in Vacutainers or CPT containing sodium heparin. HIV RNA levels were higher in plasma than in serum, regardless of collection tube or anticoagulant. Delayed centrifugation of specimens resulted in decreased HIV RNA levels, but decreases were specific to the type of tube and anticoagulant used during collection, and they were more pronounced when tubes were kept at room temperature rather than 4 degrees C. As long as centrifugation occurred immediately, plasma could be left in the collection tube for up to 30 h at 4 degrees C post centrifugation without affecting HIV RNA levels. Storage of acid-citrate-dextrose plasma in new plastic tubes (collected in Vacutainers), for 30 hours, at room temperature, after centrifugation resulted in significantly decreased HIV RNA levels compared to acid-citrate-dextrose plasma that was frozen within 2 h of collection.

Studies

Study Purpose

The purpose of this study was to determine the effects of collection tube type, anticoagulant, delayed centrifugation, and storage time and temperature on HIV RNA levels in plasma and serum. After delayed centrifugation or experimental storage, all plasma and serum was frozen at -70 degrees C prior to analysis.

Summary of Findings:

Delayed centrifugation of specimens collected into CPT resulted in significantly lower RNA levels compared to CPT specimens that were centrifuged within 2 h and left in the collection tube for up to 30 h at 4 degrees C post-centrifugation. This effect was greatest when uncentrifuged whole blood was stored in CPT at room temperature rather than 4 degrees C. The highest levels of HIV RNA were recovered when specimens were collected in either Vacutainers or CPT containing potassium EDTA. The next highest levels of HIV RNA were measured in specimens collected in acid-citrate-dextrose Vacutainers and sodium citrate CPT. The lowest levels of HIV RNA were measured in specimens collected in sodium heparin Vacutainers or CPT. HIV RNA levels were higher in plasma than in serum collected in serum separation tubes (SST), regardless of plasma collection tube or anticoagulant.

Biospecimens

Preservative Types

Frozen

Diagnoses:

AIDS/HIV-related

Platform:

Analyte	Technology Platform
RNA	Branch DNA assay

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Acquisition	Type of collection container/solution	CPT SST Standard Vacutainer
Biospecimen Acquisition	Anticoagulant	Sodium heparin Acid-citrate-dextrose Sodium citrate Potassium EDTA
Storage	Storage duration	0 h 8 h 30 h
Storage	Storage temperature	Room temperature 4 degrees C
Biospecimen Aliquots and Components	Blood and blood products	Plasma Serum
Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated

Study Purpose

The purpose of this study was to determine the effects of collection tube type, anticoagulant, and storage of plasma and plasma plus peripheral blood mononuclear cells (PBMC) at room temperature on HIV RNA levels in plasma. After experimental storage, all plasma and serum was frozen at -70 degrees C prior to analysis.

Summary of Findings:

Storage of acid-citrate-dextrose plasma (collected in Vacutainers) in new plastic tubes, for 30 hours, at room temperature, after centrifugation, resulted in significantly decreased HIV RNA levels compared to plasma that was frozen within 2 h of collection. Insignificant decreases in HIV RNA levels were observed for specimens collected in sodium citrate CPT and plasma preparation tubes (PPT) and stored at room temperature for 30 h in new tubes. Insignificant decreases in HIV RNA levels were also observed after plasma from all three tube types was stored at room temperature for 6 h.

Biospecimens

Preservative Types

Frozen

Diagnoses:

AIDS/HIV-related

Platform:

Analyte	Technology Platform
RNA	Branch DNA assay

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Acquisition	Type of collection container/solution	CPT PPT Vacutainer
Biospecimen Acquisition	Anticoagulant	Acid-citrate-dextrose Sodium citrate
Biospecimen Aliquots and Components	Blood and blood products	Plasma Plasma with peripheral blood mononuclear cells
Storage	Time at room temperature	2 h 6 h 30 h

Study Purpose

The purpose of this study was to determine the effects of collection tube type, anticoagulant, delayed centrifugation, and storage of plasma at room temperature on HIV RNA levels in plasma. After experimental storage, all plasma was frozen at -70 degrees C prior to analysis.

Summary of Findings:

Delaying centrifugation for up to 30 h and storing whole blood in the collection tube at room temperature resulted in decreased levels of HIV RNA measured in plasma for all tube types and anticoagulants. HIV RNA was the most stable and showed the smallest decrease (13%, not significant) when potassium EDTA Vacutainers were used. HIV RNA decreased by the most in sodium heparin Vacutainers, acid-citrate-dextrose Vacutainers, and PPT (34, 33, and 33% respectively).

Biospecimens

Preservative Types

Frozen

Diagnoses:

AIDS/HIV-related

Platform:

Analyte	Technology Platform
RNA	Branch DNA assay

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Acquisition	Type of collection container/solution	CPT Vacutainer PPT
Biospecimen Acquisition	Anticoagulant	Sodium heparin Potassium EDTA Acid-citrate-dextrose Sodium citrate
Biospecimen Aliquots and Components	Blood and blood products	Plasma with peripheral blood mononuclear cells Whole blood
Storage	Time at room temperature	2 h 6-8 h 30 h
Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated

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