Mole L, Margolis D, Carroll R, Todd J, Holodniy M
ARC-AIDS Research Center, Veterans Affairs Medical Center, Palo Alto, California 94304.
J Clin Microbiol. 1994 Sep;32(9):2212-5. doi: 10.1128/jcm.32.9.2212-2215.1994.
We evaluated the stability of human immunodeficiency virus (HIV) load markers from blood samples collected in VACUTAINER CPT or standard VACUTAINER brand tubes using sodium heparin or sodium citrate as anticoagulants. Quantitative plasma culture and p24 antigen concentrations were determined, and HIV RNA levels in plasma were measured by both reverse transcription-PCR-enzyme-linked immunosorbent assay (RT-PCR-ELISA) and branched DNA methods. All tubes were stored at room temperature for analysis at 2, 24, 48, and 72 h after the blood samples were drawn. No difference was seen between tube types with respect to the HIV titer in plasma or the positivity rate for all samples that demonstrated a fall in titer over time. Unbound p24 antigen levels in plasma decreased during the initial 48-h period in both tube types. Immune complex-dissociated p24 antigen levels decreased in CPT tubes but not in standard VACUTAINER tubes. The HIV RNA copy number in plasma measured by RT-PCR-ELISA was stable in most subjects and was significantly higher in CPT tubes than in standard VACUTAINER tubes at 24 and 72 h after the blood samples were drawn. The branched DNA probe assay detected a significant decline in HIV RNA equivalent in plasma over 72 h in both collection tubes, the decline being more dramatic in the standard VACUTAINER tube than the CPT tube. Overall, interday variability suggests that samples collected for a particular assay should be processed at the same time after blood is drawn and that a particular tube type be used throughout a given study.
我们评估了使用肝素钠或枸橼酸钠作为抗凝剂,在VACUTAINER CPT管或标准VACUTAINER品牌管中采集的血液样本中人类免疫缺陷病毒(HIV)载量标志物的稳定性。测定了定量血浆培养物和p24抗原浓度,并通过逆转录-聚合酶链反应-酶联免疫吸附测定(RT-PCR-ELISA)和分支DNA方法测量了血浆中的HIV RNA水平。所有试管均在室温下保存,以便在采血后2、24、48和72小时进行分析。就血浆中的HIV滴度或所有随时间滴度下降的样本的阳性率而言,不同试管类型之间未见差异。两种试管类型中,血浆中未结合的p24抗原水平在最初48小时内均下降。CPT管中免疫复合物解离的p24抗原水平下降,而标准VACUTAINER管中则未下降。通过RT-PCR-ELISA测量的血浆中HIV RNA拷贝数在大多数受试者中是稳定的,并且在采血后24和72小时,CPT管中的该拷贝数显著高于标准VACUTAINER管。分支DNA探针测定法检测到两种采集管中血浆中的HIV RNA等效物在72小时内均显著下降,标准VACUTAINER管中的下降比CPT管更显著。总体而言,日间变异性表明,为特定检测采集的样本应在采血后同时进行处理,并且在给定研究中应始终使用特定的试管类型。