Automation of nucleic acid extraction for NAT screening of individual blood units.
Automation of nucleic acid extraction for NAT screening of individual blood units.
复制标题
自动化核酸提取，用于单个血液单位的 NAT 筛查。
DOI：
10.1046/j.1537-2995.2001.41040483.x
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发表时间：
2001
期刊：
影响因子：
2.9
通讯作者：
Prince,AM
中科院分区：
文献类型：
--
作者：
Lee,DH;Prince,AM
背景：在扩增程序之前从样品中提取核酸所涉及的劳动密集型步骤目前阻碍了单单位血液NAT的自动化。一种新的方法已经开发了这些步骤的自动化使用亲水性聚偏氟乙烯（PVDF）filterplate.Study设计和方法：定量核酸回收率从血清中含有HCV，HIV，HBV，HAV，和人细小病毒B19和从3H标记的HCV RNA平行测定半自动PVDF方法和单柱方法（Qiagen）。结果：PVDF法和Qiagen单柱法的HCV、HIV和HBV（硅胶珠）回收率相似。在连续稀释的血清样本中，基于PVDF的PCR检测试剂盒对HCV、HIV和HBV的灵敏度始终在相同检测试剂盒中使用Qiagen单柱方法时获得的两个连续稀释度范围内。与使用of3H标记的HCV RNA，回收率约70%，发现由两种methods.CONCLUSION：PVDF方法将允许完全自动化的同时提取核酸从血清中含有HCV，HIV和HBV。该程序将允许对单个血液单位进行NAT筛查，将取代目前的合并筛查，并将在减少劳动力和成本的情况下提高血液安全性。
BACKGROUND:Automation of NAT for single units of blood is currently hampered by the labor‐intensive steps involved in the extraction of nucleic acids from samples before the amplification procedures. A new method has been developed for the automation of these steps using hydrophilic polyvinylidene fluoride (PVDF) filter plates.STUDY DESIGN AND METHODS:Quantitative nucleic acid recoveries from sera containing HCV, HIV, HBV, HAV, and human parvovirus B19 and from3H‐labeled HCV RNA were determined in parallel by the semi‐automated PVDF method and a single‐column method (Qiagen). Quantitative PCR was performed.RESULTS:Similar recoveries of HCV, HIV, and HBV (with silica beads) were observed with the PVDF method and with the Qiagen single‐column method. The sensitivity of the PVDF‐based PCR assay for HCV, HIV, and HBV in serially diluted serum samples was always within two serial dilutions of that obtained when the Qiagen single‐column method was used in the same assays. With the use of3H‐labeled HCV RNA, recoveries of approximately 70 percent were found by both methods.CONCLUSION:The PVDF method will permit full automation of the simultaneous extraction of nucleic acid from sera containing HCV, HIV, and HBV. This procedure will permit NAT screening of individual units of blood, will replace the current screening of pools, and will achieve improved blood safety with reduced labor and costs.