Bruisten S M, Oudshoorn P, van Swieten P, Boeser-Nunnink B, van Aarle P, Tondreau S P, Cuypers H T
Central Laboratory, Netherlands Red Cross Blood Transfusion Service, Department of Viral Diagnostics, Amsterdam.
J Virol Methods. 1997 Sep;67(2):199-207. doi: 10.1016/s0166-0934(97)00097-9.
The influence of different storage temperatures and anticoagulation conditions on the HIV-1 RNA load as measured by NASBA-QT was examined. Blood specimens from 14 HIV-1 infected individuals were processed within 2 h after collection. The HIV-1 RNA load remained stable for at least 6 months when samples were frozen directly at -70 degrees C in lysis buffer. This lysis buffer fully inactivated the virus. When whole EDTA blood was stored, the HIV-1 RNA load was stable for 72 h at 25 degrees C, but it declined within 24 h at 4 degrees C. The HIV-1 RNA load in whole heparinized blood declined significantly after 24 h at both 4 and 25 degrees C. It was slightly lower (average of 0.18 log ml-1) than in whole EDTA blood. At 4 degrees C, the HIV-RNA load in serum and EDTA-plasma stored with lysis buffer did not decline up to 14 days. At + 30 degrees C, however, the load declined significantly after 2 days. Of clinical significance, the mean load in EDTA plasma was 0.5 log ml-1 higher than in serum. This difference was patient dependent (range 0.1-0.7 log ml-1). We thus recommend, for quantifying HIV-1 RNA by NASBA, to use preferably EDTA blood which is kept at room temperature until plasma separation. When using heparinized blood, the plasma should be stored frozen within 8 h.
研究了不同储存温度和抗凝条件对通过NASBA-QT检测的HIV-1 RNA载量的影响。14名HIV-1感染者的血液标本在采集后2小时内进行处理。当样本在裂解缓冲液中直接于-70℃冷冻时，HIV-1 RNA载量至少6个月保持稳定。这种裂解缓冲液可使病毒完全失活。当储存全血乙二胺四乙酸（EDTA）时，HIV-1 RNA载量在25℃可稳定72小时，但在4℃时24小时内就会下降。全血肝素化后，在4℃和25℃下24小时后HIV-1 RNA载量均显著下降。其略低于全血EDTA中的载量（平均低0.18 log ml-1）。在4℃时，用裂解缓冲液储存的血清和EDTA血浆中的HIV-RNA载量在14天内均未下降。然而，在30℃时，2天后载量显著下降。具有临床意义的是，EDTA血浆中的平均载量比血清中的高0.5 log ml-1。这种差异因患者而异（范围为0.1 - 0.7 log ml-1）。因此，我们建议，为通过NASBA定量检测HIV-1 RNA，最好使用EDTA血液，并在室温下保存直至分离血浆。当使用肝素化血液时，血浆应在8小时内冷冻保存。
