Effects of handling and storage of blood on the stability of hepatitis C virus RNA: implications for NAT testing in transfusion practice.
OBJECTIVE
To determine the stability of hepatitis C virus (HCV) RNA during transport and storage of blood samples from donors, prior to screening for HCV by nucleic acid amplification technology.


METHODS
Various blood and plasma sample types were stored for up to 120 h at different temperatures and the HCV RNA level was measured using an in house quantitative reverse transcription-polymerase chain reaction.


RESULTS
No decline in HCV RNA level was observed after 72 h of storage of whole blood at 4 degrees C in EDTA tubes (Greiner) and Plasma Preparation Tubes (PPT; Becton Dickinson), while insignificant declines of 0.2 log10 and 0. 25 log10 occurred at 25 degrees C after 72 h in the EDTA tubes and PPT tubes, respectively. When whole blood was stored with mixed anticoagulants CPDA-1 and EDTA for up to 120 h, no decline in HCV RNA level was observed at 4 degrees C and 25 degrees C, while a significant decline of 0.37 log10 occurred at 37 degrees C after 120 h. The temperature during transportation was investigated with a 12-hour period at 25 degrees C and 37 degrees C before storage at 4 degrees C for 108 h. Neither temperature resulted in any loss of HCV RNA in comparison with 120 h of storage at 4 degrees C.


CONCLUSIONS
Whole blood anticoagulated with EDTA or CPDA-1/EDTA may be stored at up to 25 degrees C (room temperature) for up to 5 days without any significant loss in plasma HCV RNA level.
Grant
P R
PR
Royal Free and University College Medical School, Department of Virology, London, UK. paul.grant@ucl.ac.uk
Kitchen
A
A
Barbara
J A
JA
Hewitt
P
P
Sims
C M
CM
Garson
J A
JA
Tedder
R S
RS
D000225:Adenine / Q000494:pharmacology; D000925:Anticoagulants / Q000494:pharmacology; D001793:Blood Preservation / Q000009:adverse effects* / Q000592:standards; D001803:Blood Transfusion; D002614:Chelating Agents / Q000494:pharmacology; D002951:Citrates / Q000494:pharmacology; D003451:Cryoprotective Agents / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004739:England; D005784:Gene Amplification; D005947:Glucose / Q000494:pharmacology; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D008403:Mass Screening; D010710:Phosphates / Q000494:pharmacology; D010949:Plasma / Q000821:virology; D019064:Product Packaging; D012367:RNA, Viral / Q000097:blood* / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013048:Specimen Handling / Q000592:standards*; D013696:Temperature; D013997:Time Factors
D000925:Anticoagulants; C034876:CPDA solutions; D002614:Chelating Agents; D002951:Citrates; D003451:Cryoprotective Agents; D010710:Phosphates; D012367:RNA, Viral; D004492:Edetic Acid; D005947:Glucose; D000225:Adenine