Sankary T M, Yang G, Romeo J M, Ulrich P P, Busch M P, Rawal B D, Vyas G N
Department of Laboratory Medicine, University of California, San Francisco.
Transfusion. 1994 Aug;34(8):656-60. doi: 10.1046/j.1537-2995.1994.34894353458.x.
Since screening for antibody to hepatitis C virus (HCV) was introduced in 1990, posttransfusion hepatitis has been reduced to nearly background levels. This has led to reconsideration of the value of testing donated blood for elevated alanine aminotransferase (ALT). The contribution of ALT testing in detecting seronegative infection was evaluated by the performance of polymerase chain reaction (PCR) for hepatitis B virus (HBV) or HCV in plasma from ALT-elevated blood units.
Testing was performed on 375 units of plasma, derived from an equivalent of 47,500 blood donations, with a highly sensitive hemi-nested PCR procedure. Using a triplet of primers directed at the conserved regions of HBV DNA and 5'-noncoding regions of HCV RNA, the hemi-nested PCR assay can reliably amplify 10 viral molecules to levels detectable in ethidium bromide-stained agarose gels. Pools of plasma from groups of four donors were screened with hemi-nested PCR. For any reactive pools, the plasma from individual donors was retested twice on different aliquots.
Two of 375 units, both with midrange ALT elevation, were repeatedly reactive in hemi-nested PCR (one each for HBV DNA and HCV RNA). However, samples from the two suspect donors tested 9 and 5 months later revealed no seroconversion, elevated ALT, or viral genomes in hemi-nested PCR.
The lack of confirmed HBV or HCV infection in this study representing an estimated 47,500 voluntary blood donations suggests that routine ALT testing for further prevention of posttransfusion hepatitis after exclusion of HBV- and/or HCV-seropositive blood may be superfluous.
自1990年引入丙型肝炎病毒(HCV)抗体筛查以来,输血后肝炎已降至接近背景水平。这导致人们重新审视检测献血者丙氨酸转氨酶(ALT)升高的价值。通过对ALT升高的血液单位血浆中乙型肝炎病毒(HBV)或HCV进行聚合酶链反应(PCR),评估了ALT检测在检测血清阴性感染中的作用。
使用高度敏感的半巢式PCR程序对相当于47500次献血的375单位血浆进行检测。半巢式PCR检测使用针对HBV DNA保守区域和HCV RNA 5'非编码区域的三联体引物,可将10个病毒分子可靠地扩增到溴化乙锭染色琼脂糖凝胶中可检测的水平。对来自四名献血者一组的血浆池进行半巢式PCR筛查。对于任何反应性血浆池,对个体献血者的血浆在不同等分试样上进行两次重新检测。
375单位血浆中有两单位,ALT均为中度升高,在半巢式PCR中反复呈反应性(分别为HBV DNA和HCV RNA各一单位)。然而,9个月和5个月后对两名可疑献血者的样本检测显示,半巢式PCR中未出现血清转化、ALT升高或病毒基因组。
本研究中代表约47500次自愿献血的样本未确诊HBV或HCV感染,这表明在排除HBV和/或HCV血清阳性血液后,为进一步预防输血后肝炎进行常规ALT检测可能是多余的。