输血相容性检测室内质控品制备技术优化与性能评价

Performance evaluation and optimization of preparation technology of internal quality control products for blood transfusion compatibility testing

目的优化自制输血相容性检测室内全血质控品制备技术,检测其保存过程中综合性能指标的变化,确定合理的处理流程、保存期限并评价其应用价值。方法选择多人份采集时间≤10 d的B型RhD阴性健康献血者标本,将其混合后离心留取上清血浆,再将混合浓缩红细胞分成2组,MAP组:使用MAP红细胞保养液洗涤2次;Saline组:使用生理盐水洗涤2次。将2组洗涤后的红细胞分别与MAP红细胞保存液、对应混合血浆按照1∶2∶3的体积比混合。选择商品化IgG抗-D试剂做抗-D效价测定,确定出现最后1个2+凝集强度的稀释倍数,并按照此稀释倍数分别在2组质控品中填加相应体积的IgG抗-D。将2组混合悬液分装在硬质塑料试管中盖帽4℃保存,每天在室温放置1 h,分别在保存的0、35、42、49 d检测质控品标本红细胞与标准抗-B的凝集强度、IgM抗-A与反定A细胞的凝集强度、IgG抗-D与RhD阳性O型红细胞的凝集强度、上清液中Na+、K+、LDH、乳酸、FHb浓度、红细胞形态变化以及质控品中细菌繁殖情况。结果保存过程中2组质控品红细胞B抗原、IgG抗D抗体反应活性均无明显变化(P>0.05),IgM抗A抗体反应活性虽出现波动(P<0.01),但凝集强度变化均在1+范围内;2组质控品K+、乳酸浓度均随保存时间延长明显增高(P<0.01),但保存35、42d时2组各指标之间无明显差异(P>0.05);2组质控品FHb浓度均随保存时间延长而增高,但相同保存时间2组之间比较无明显差异(P<0.01)。保存49 d时MAP组和Saline组FHb浓度分别为(857.1±301.5)mg/L和(595.3±334.9)mg/L,与保存0d相比均明显升高(P<0.01),MAP组部分质控品FHb浓度已经超过中度溶血标准(1 000 mg/L);2组质控品保存末期(≤42 d)红细胞都会出现一定程度的皱缩并形成棘突,但2组之间无明显差异;2组质控品保存过程中都未见细菌生长。结论本室采用2种方法自制的全血质控品质量无明显差异,保存期都能达到42 d,且管间差异小、抗原抗体反应活性稳定,能够满足输血相容性检测室内质控的相关要求,适合在输血相容性检测实验室推广。

Objective To optimize the technology for preparing internal quality control（IQC） products for blood transfusion compatibility testing,and detect changes in performance indicators of IQC products during storage,and determine a reasonable processing and shelf life,and evaluate the application value of prepared IQC products.Methods B/RhD-healthy blood donor samples within 10 days of collection were mixed and centrifuged and prepared as mixed packed red blood cells and mixed plasma.Mixed packed red blood cells were divided into two groups,and one group was washed twice by using the MAP RBC preservative solution（MAP group） and the other group was washed twice with normal saline（Saline group）.Washed red blood cells from the two groups,MAP red blood cell preservation solution,and the mixed plasma were mixed at 1∶2∶3 by volume.Antibody titer of IgG anti-D reagent was deteceted,and the highest dilution resulting in 2＋ agglutination strength was determined.According to the dilution determined,IgG anti-D reagent was added to the above red cell,MAP and plasma mixture.The two groups of mixed suspension were aliquoted in capped rigid plastic tubes and stored at 4℃.All quality control samples were placed in room temperature for 1h every day.B antigen on the red blood cells,IgM anti-A antibody,IgG anti-D antibody,Na＋,K＋,LDH,lactate and free hemoglobin（FHb） concentration in the supernatant were detected on 0,35,42,49 days of storage.Results The reactivity of B antigen and IgG anti-D antibody in two groups did not change significantly（P 0.05）.There were fluctuations in activity of IgM anti-A antibody,but the changes of agglutination intensity were in the acceptable 1＋ range.K＋ and lactate concentration in the IQC products of the two groups significantly increased with prolonged storage time（P0.01）,but there were no significant differences between two groups on 35,42 days of storage（P0.05）.FHb concentration in the IQC products of the two groups increased with the extension of storage time,there was no significant difference between the two groups at the same time of storage（P0.05）.FHb concentration in MAP group and Saline group on 49 days of storage were（857.1 ± 301.5） mg / L and（595.3 ± 334.9） mg / L respectively,and significantly increased compared with those on 0 day（P0.01）.FHb concentration in some IQC products from MAP group exceeded standard concentration of moderate hemolysis（1 000 mg / L）.Although shrinkage and spinous process appeared on a certain proportion of red blood cells in two groups on 35,42 days of storage,there was no significant difference between the two groups.There was no bacterial growth in the IQC products from two groups.Conclusion The IQC products of recomposed whole blood prepared by two methods can be preserved effectively for at least 42 days,and,the small tube-to-tube variation and the good stability of antigen-antibody reactivity can meet IQC requirements for blood transfusion compatibility testing.