Stability of hepatitis C virus RNA during specimen handling and storage prior to NASBA amplification

The influence of different anticoagulants and pre-amplification storage conditions on the stability of hepatitis C virus (HCV)-RNA, as detected by the quantitative HCV NASBA assay (NASBA-QT), was studied. The HCV-RNA load remained stable for at least 15 months when serum or plasma samples (EDTA and...

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Bibliographic Details
Published in:Journal of virological methods 1998-06, Vol.72 (2), p.175-184
Main Authors: Damen, M, Sillekens, P, Sjerps, M, Melsert, R, Frantzen, I, Reesink, H.W, Lelie, P.N, Cuypers, H.T.M
Format: Article
Language:English
Subjects:
Biological and medical sciences
Buffers
Edetic Acid - pharmacology
Fundamental and applied biological sciences. Psychology
Gene Amplification
HCV-RNA
Hepacivirus - drug effects
Hepacivirus - genetics
Heparin - pharmacology
Humans
Lysis buffer
Microbiology
NASBA-QT
RNA, Viral - blood
RNA, Viral - drug effects
RNA, Viral - isolation & purification
Specimen Handling
Stability
Storage conditions
Techniques used in virology
Temperature
Time Factors
Virology
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Summary:The influence of different anticoagulants and pre-amplification storage conditions on the stability of hepatitis C virus (HCV)-RNA, as detected by the quantitative HCV NASBA assay (NASBA-QT), was studied. The HCV-RNA load remained stable for at least 15 months when serum or plasma samples (EDTA and heparin) were directly frozen at −70°C in lysis buffer. At 4°C, the HCV-RNA load in serum or plasma stored with lysis buffer did not decline for at least 14 days. At 30°C, however, the load declined significantly after 7 days. When clotted, whole blood was stored at 4°C, the HCV-RNA load was stable for 72 h. However, when EDTA-anticoagulated whole blood was stored at 4°C, the HCV-RNA load declined significantly after 48 h. In paired plasma and serum samples at baseline the HCV-RNA levels were similar. Heparin did not influence the efficiency of the HCV NASBA-QT assay. Clotted blood as well as EDTA or heparin anticoagulated blood can be used for quantifying HCV-RNA using the NASBA-QT assay. Blood samples should be stored at 4°C after collection and serum or plasma separated within 24 h. Preferably, after separation, samples should be frozen in lysis buffer at −70°C until NASBA-QT analysis.
ISSN:0166-0934
1879-0984
DOI:10.1016/S0166-0934(98)00024-X