QUANTITATIVE DETECTION OF HEPATITIS-C VIRUS-RNA WITH A SOLID-PHASE SIGNAL AMPLIFICATION METHOD - DEFINITION OF OPTIMAL CONDITIONS FOR SPECIMEN COLLECTION AND CLINICAL-APPLICATION IN INTERFERON-TREATED PATIENTS
QUANTITATIVE DETECTION OF HEPATITIS-C VIRUS-RNA WITH A SOLID-PHASE SIGNAL AMPLIFICATION METHOD - DEFINITION OF OPTIMAL CONDITIONS FOR SPECIMEN COLLECTION AND CLINICAL-APPLICATION IN INTERFERON-TREATED PATIENTS
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DOI：
10.1002/hep.1840190603
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发表时间：
1994-06-01
期刊：
影响因子：
13.5
通讯作者：
ALBRECHT, J
中科院分区：
文献类型：
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作者：
DAVIS, GL;LAU, JYN;ALBRECHT, J
为了确定用于定量丙型肝炎病毒RNA测定的血清标本制备的最佳条件，对患者样本进行处理，以便可以独立评估从凝块形成到离心、离心到分离血清和收集血清直至冷冻的时间差异。还测定了多次冻融循环的影响。当从凝块形成到离心分离的时间超过2小时时，丙型肝炎病毒RNA活性会逐渐显著降低。6小时后降低32%，24小时后降低49%。如果在凝块形成后立即进行离心分离，即使血清与凝块在6小时内未分离，丙型肝炎病毒RNA活性的损失也降至10%以下。（血清分离管）防止丙型肝炎病毒RNA活性损失长达24小时。丙型肝炎病毒RNA活性没有损失多达三个冻融循环。当在这些最佳条件下制备患者标本时，定量分支DNA信号扩增试验在丙型肝炎病毒感染患者中的灵敏度为83%，在肝病患者中的特异性为100%。丙型肝炎病毒RNA水平的波动显示与重组干扰素-α治疗患者中观察到的生化变化相关（2b）。这些数据表明，不正确或不一致的血清制备方法可能导致丙型肝炎病毒RNA水平假性降低和不可靠。丙型肝炎病毒RNA定量检测的临床应用只有在血清制备得当时才有可能。
To determine the optimal conditions for preparation of serum specimens for quantitative hepatitis C virus RNA determination, patient samples were processed such that differences in time from clot formation to centrifugation, centrifugation to separation of serum and collection of serum until freezing could be independently assessed. The effects of multiple cycles of freezing and thawing were also determined. There was progressive and significant loss of hepatitis C virus RNA activity when the time from the formation of the clot until centrifugation was longer than 2 hr. This reduction reached 32% after 6 hr and 49% after 24 hr. if centrifugation was performed immediately after formation of the clot, loss of hepatitis C virus RNA activity was reduced to less than 10% even though the serum remained unseparated from the clot for up to 6 hr. Centrifugation of blood through a paraffin plug (serum separator tube) prevented loss of hepatitis C virus RNA activity for up to 24 hr. There was no loss of hepatitis C virus RNA activity with up to three freeze-thaw cycles. When patient specimens were prepared under these optimal conditions, the sensitivity of the quantitative branched DNA signal amplification assay in patients with hepatitis C virus infection was 83% and the specificity in patients with liver disease was 100%. Fluctuations in hepatitis C virus RNA levels were shown to correlate with biochemical changes observed in patients treated with recombinant interferon-alpha(2b). These data demonstrate that improper or inconsistent methods of serum preparation may result in falsely low and unreliable levels of hepatitis C virus RNA. Clinical applications of quantitative hepatitis C virus RNA assays will only be possible when sera are properly prepared.