Laden F, Wolff M S, Niguidula N J, Spiegelman D, Hankinson S E, Hunter D J
Department of Epidemiology, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Cancer Epidemiol Biomarkers Prev. 1997 May;6(5):333-8.
Because archived blood specimens are an important but limited resource for conducting epidemiological studies using biomarkers, it is important to develop analytical techniques that minimize the amount of sample needed. We modified an established 1.0-ml blood plasma organochlorine assay to use smaller volumes. We assessed its utility by comparing the accuracy and precision of measurements obtained with different-sized aliquots of spiked plasma from three pools of known concentration of 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) and polychlorinated biphenyls (PCBs; low, medium, and high). There was a modest sacrifice in accuracy using 0.5 as opposed to 1.0 ml. However, the within-batch coefficients of variation, a measure of laboratory error, were consistently low when 0.5-ml aliquots were used. For both DDE and PCB concentrations, this error was less than 5% for the medium and high pools [5-20 parts per billion (ng/ml)] and less than 9% for the low pool (< 1 part per billion). After determining that aliquots of 0.5 ml were sufficient, we performed a blinded quality control analysis of stored plasma. In this study, the within-subject variation was low for DDE and PCBs and substantially lower than the between-subject variation, suggesting that the assay would rank subjects with reasonable precision. Our results suggest that use of 0.5-ml as opposed to 1.0-ml aliquots should not compromise the power of a nested case-control study to detect differences between subjects and would thus save plasma for future research. For populations with very low levels of organochlorines, however, the larger volumes should still be used.
由于存档血液样本是使用生物标志物进行流行病学研究的重要但有限的资源,因此开发能将所需样本量降至最低的分析技术非常重要。我们对已有的1.0毫升血浆有机氯检测方法进行了改进,以使用更小的体积。我们通过比较从三个已知浓度的1,1-二氯-2,2-双(对氯苯基)乙烯(DDE)和多氯联苯(PCBs;低、中、高浓度)混合样本中取出的不同体积加标血浆所获得测量值的准确性和精密度,来评估其效用。与使用1.0毫升相比,使用0.5毫升时准确性略有牺牲。然而,当使用0.5毫升等分样本时,批内变异系数(衡量实验室误差的指标)一直很低。对于DDE和PCB浓度,中高浓度混合样本(5 - 20纳克/毫升)的误差小于5%,低浓度混合样本(< 1纳克/毫升)的误差小于9%。在确定0.5毫升等分样本足够后,我们对储存血浆进行了盲法质量控制分析。在这项研究中,DDE和PCBs的个体内变异较低,且远低于个体间变异,这表明该检测方法能够以合理的精度对受试者进行排名。我们的结果表明,使用0.5毫升而非1.0毫升等分样本不会影响巢式病例对照研究检测受试者之间差异的能力,因此可为未来研究节省血浆。然而,对于有机氯水平非常低的人群,仍应使用较大体积的样本。