Lelie P Nico, van Drimmelen Harry A J, Cuypers H Theo M, Best Susan J, Stramer Susan L, Hyland Catherine, Allain Jean-Pierre, Moncharmont Pierre, Defer Christine, Nübling Micha, Glauser Andreas, da Silva Cardoso Marcia, Viret Jean-François, Lankinen Mervi H, Grillner Lena, Wirthmüller Urs, Coste Joliette, Schottstedt Volkmar, Masecar Barbara, Dax Elizabeth M
VQC Laboratory, Sanquin-CLB Diagnostic Division, Amsterdam, The Netherlands.
Transfusion. 2002 May;42(5):527-36. doi: 10.1046/j.1537-2995.2002.00101.x.
The FDA requirement for sensitivity of viral NAT methods used in blood screening is a 95-percent detection limit of 100 copies per mL, whereas the NAT screening system should have a sensitivity of at least 5000 copies per mL per individual donation. According to the Common Technical Specifications of the European Directive 98/79/EC for in vitro diagnostics, viral standard dilutions (calibrated against the WHO standard) should be tested at least 24 times for a statistically valid assessment of the 95-percent detection limit.
Viral standard dilution panels (PeliCheck, VQC-CLB) were prepared for HCV RNA genotypes 1 and 3 and for HIV RNA genotypes B and E. In a multicenter study, 23 laboratories tested the panels all together in 8 to 91 test runs per NAT method.
The following 95-percent detection limits (and 95% CIs) were found on the HCV RNA genotype 1 reference panels (shown as geq/mL): Gen-Probe TMA, 85 (64-118); AmpliScreen, 126 (83-225); AmpliScreen with NucliSens Extractor, 21 (13-44); Amplicor with NucliSens Extractor, 69 (50-102), and Amplicor with Qiagen extraction technology, 144 (74-102). On HIV RNA genotype B dilution panels, the following 95-percent detection limits were found (shown as geq/mL): Gen-Probe TMA, 31 (20-52); AmpliScreen, 126 (67-311); AmpliScreen with NucliSens Extractor, 37 (23-69), and NucliSens QL assay, 123 (51-566). HIV RNA genotype E panels were detected with equal sensitivity as HIV RNA genotype B panels. In the Gen-Probe TMA assay, the 50-percent detection limits on HIV RNA type B and type E were 3.6 (2.6-5.0) and 3.9 (2.4-5.8) geq per mL, respectively. The HCV RNA genotype 1 and 3 standards were detected with equal sensitivity.
The differences in sensitivity between NAT assays can be explained by the input of isolated viral nucleic acid in the amplification reactions. The FDA requirements for sensitivity of NAT blood screening assays can be met by the Gen-probe TMA, as well as by the AmpliScreen assays, particularly when combined with the NucliSens Extractor.
美国食品药品监督管理局（FDA）对用于血液筛查的病毒核酸扩增检测（NAT）方法的灵敏度要求是，95%检测限为每毫升100拷贝，而NAT筛查系统对每份个体献血的灵敏度应至少为每毫升5000拷贝。根据欧洲指令98/79/EC关于体外诊断的通用技术规范，病毒标准稀释液（根据世界卫生组织标准校准）应至少检测24次，以便对95%检测限进行统计学上有效的评估。
针对丙型肝炎病毒（HCV）RNA 1型和3型以及人类免疫缺陷病毒（HIV）RNA B型和E型制备了病毒标准稀释液板（PeliCheck，VQC-CLB）。在一项多中心研究中，23个实验室对这些稀释液板进行了检测，每种NAT方法共进行了8至91次测试。
在HCV RNA 1型参考稀释液板上发现以下95%检测限（及95%置信区间）（以每毫升基因组当量[geq/mL]表示）：Gen-Probe转录介导扩增法（TMA），85（64 - 118）；AmpliScreen，126（83 - 225）；搭配NucliSens提取仪的AmpliScreen，21（13 - 44）；搭配NucliSens提取仪的Amplicor，69（50 - 102），以及搭配Qiagen提取技术的Amplicor，144（74 - 202）。在HIV RNA B型稀释液板上，发现以下95%检测限（以geq/mL表示）：Gen-Probe TMA，31（20 - 52）；AmpliScreen，126（67 - 311）；搭配NucliSens提取仪的AmpliScreen，37（23 - 69），以及NucliSens QL检测法，123（51 - 566）。HIV RNA E型稀释液板的检测灵敏度与HIV RNA B型稀释液板相同。在Gen-Probe TMA检测法中，HIV RNA B型和E型的50%检测限分别为每毫升3.6（2.6 - 5.0）和3.9（2.4 - 5.8）geq。HCV RNA 1型和3型标准品的检测灵敏度相同。
NAT检测方法之间灵敏度的差异可以通过扩增反应中分离的病毒核酸输入量来解释。Gen-probe TMA以及AmpliScreen检测方法，特别是与NucliSens提取仪联合使用时，能够满足FDA对NAT血液筛查检测方法灵敏度的要求。
