Wakamatsu C, Takakura F, Kojima E, Kiriyama Y, Goto N, Matsumoto K, Oyama M, Sato H, Okochi K, Maeda Y
Fukuoka Red Cross Blood Center, Fukuoka, Japan.
Vox Sang. 1999;76(1):14-21. doi: 10.1159/000031014.
Human parvovirus B19 (B19 virus) can be transmitted through blood transfusion and plasma-derived products. In a previous report, we utilized the simple hemagglutination method based on the interaction between the B19 virus and P antigen on human erythrocytes in order to screen the blood donors. We called this method receptor-mediated hemagglutination (RHA) [Lancet 1995;346:1237-1238]. In this paper, we report on a large-scale screening of the B19 virus by RHA and discuss the results.
Donor sera from September 1995 to March 1997 and seroconversion panels were enrolled. Donor sera were examined by RHA for large-scale screening. The positive sera in the first screening were then further investigated by the RHA inhibition test, countercurrent immunoelectrophoresis (CIE), an enzyme-linked immunosorbent assay, and polymerase chain reaction (PCR). We also evaluated the infectivity and neutralizing activity of various kinds of sera by the erythroid colony forming unit (CFU-e) assay. To examine the detection limits of the B19 virus by RHA, B19-viremic sera were purified by sucrose gradient ultracentrifugation.
Among 257,710 sera specimens, 293 sera (0.11%) gave a positive reaction in the first screening using RHA. Out of these 293 sera specimens, 31 were positive for PCR, of which 28 were also RHA inhibition-positive, and 25 of the 28 CIE-positive. In the CFU-e injury assay, all the RHA inhibition (+) sera showed a decrease in the number of erythroid colonies. The RHA inhibition (-) PCR (+) B19 antibody (+) sera did not affect the erythroid colony formation and protected CFU-e from injury by the B19 virus. By measuring the amount of purified B19 protein and its RHA titer, the detection limit of the B19 virus by RHA was calculated to the 0.37+/-0.03 ng/ml.
These results suggest that the RHA(+) RHA inhibition (+) sera were infectious in vitro. The combination of RHA and the RHA inhibition test is considered to be useful for the large-scale screening of infectious B19 virus in blood donors with high specificity.
人细小病毒B19（B19病毒）可通过输血和血浆衍生制品传播。在之前的一份报告中，我们利用基于B19病毒与人红细胞上P抗原相互作用的简单血凝方法来筛查献血者。我们将此方法称为受体介导的血凝反应（RHA）[《柳叶刀》1995年；346：1237 - 1238]。在本文中，我们报告了通过RHA对B19病毒进行的大规模筛查并讨论结果。
纳入1995年9月至1997年3月的献血者血清以及血清转换样本。通过RHA对献血者血清进行大规模筛查。首次筛查中的阳性血清随后通过RHA抑制试验、对流免疫电泳（CIE）、酶联免疫吸附测定和聚合酶链反应（PCR）进一步检测。我们还通过红系集落形成单位（CFU - e）测定评估了各类血清的感染性和中和活性。为检测RHA对B19病毒的检测限，通过蔗糖梯度超速离心法纯化B19病毒血症血清。
在257,710份血清样本中，293份血清（0.11%）在首次使用RHA筛查时呈阳性反应。在这293份血清样本中，31份PCR呈阳性，其中28份RHA抑制试验也呈阳性，28份CIE阳性样本中有25份。在CFU - e损伤测定中，所有RHA抑制试验（+）血清的红系集落数量均减少。RHA抑制试验（ - ）PCR（+）B19抗体（+）血清不影响红系集落形成，并保护CFU - e免受B19病毒损伤。通过测量纯化的B19蛋白量及其RHA效价，计算出RHA对B19病毒的检测限为0.37±0.03 ng/ml。
这些结果表明，RHA（+）RHA抑制试验（+）血清在体外具有感染性。RHA与RHA抑制试验相结合被认为对大规模筛查具有高特异性的献血者中感染性B19病毒很有用。
