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 heparin) were directly frozen at -70 degrees C in lysis buffer. At 4 degrees C, the HCV-RNA load in serum or plasma stored with lysis buffer did not decline for at least 14 days. At 30 degrees C, however, the load declined significantly after 7 days. When clotted, whole blood was stored at 4 degrees C, the HCV-RNA load was stable for 72 h. However, when EDTA-anticoagulated whole blood was stored at 4 degrees 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 degrees C after collection and serum or plasma separated within 24 h. Preferably, after separation, samples should be frozen in lysis buffer at -70 degrees C until NASBA-QT analysis.
Viral Diagnostic Department, Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
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Damen
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Sillekens
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Sjerps
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Melsert
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Frantzen
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Reesink
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Lelie
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Cuypers
D002021:Buffers; D004492:Edetic Acid / Q000494:pharmacology*; D005784:Gene Amplification; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D012367:RNA, Viral / Q000097:blood / Q000187:drug effects* / Q000302:isolation & purification; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors
D002021:Buffers; D012367:RNA, Viral; D006493:Heparin; D004492:Edetic Acid