Grabarczyk Piotr, Korzeniowska Jolanta, Liszewski Grzegorz, Kalińska Aleksandra, Sulkowska Ewa, Krug-Janiak Maria, Kopacz Aneta, Łetowska Magdalena, Brojer Ewa
Zaklad Wirusologii, Instytut Hematologii i Transfuzjologii w Warszawie, Regionalne Centrum Krwiodawstwa i Krwiolecznictwa w Lublinie.
Przegl Epidemiol. 2012;66(1):7-12.
Since 2004 Polish blood donors have been tested for parvovirus B19 (B19V) DNA. The screening testing has been performed in donors of plasma for fractionation and anti-D and anti-HBs production and donors of erythrocytes used for immunization. AIM is to present methods of the testing, quality control and results in period 2004-2010.
Testing was performed in individual donation testing (IDT) in Regional Blood Transfusion Center (RBTC) in Lublin or in pools of 24 in Institute of Haematology and Transfusion Medicine in Warsaw (IHTM). Quantitative testing with real-time PCR was preceded with nucleic acid isolation on silica based methods (Prepito Viral DNA/RNA, Chemagen and QIAamp DNA Mini Kit, QIAGEN). Amplification was performed initially with home made method and later with commercial assay (Artus Parvo B19 RG PCR Kit on Rotor Gene 6 000). In total 17 625 donations were tested: 8 539 in pools and 9 090 individually. Beside routine external quality control programmes in which both laboratories participated (Proficiency Study VQC,Amsterdam, Holand; EQA Programe, Glasgow, Scotland), panel containing negative samples, positive with very high DNA B 19V level and plasma infected with genotype 2 was prepared for RBTC in Lublin.
B19V infection frequency was 1:980 donations, low viraemic donations were detected most frequently (1:1 037). It was identified only one donation with DNA load that could cause potential health risk for plasma product recipients (1:17 625). In one of the donors B 19V DNA was observed for 3 years and 3 months. In acute or persistent phase of infection no clinical or laboratory symptoms (morphology of peripheral blood, ALT) were observed. Due to risk of underestimation of viral load connected with viral genome polymorphism all donations with B19V positive result were not allowed to be clinically used.
自2004年起,对波兰献血者进行了细小病毒B19(B19V)DNA检测。筛查检测在用于分馏的血浆捐献者、抗-D和抗-HBs生产的血浆捐献者以及用于免疫的红细胞捐献者中进行。目的是介绍2004 - 2010年期间的检测方法、质量控制和结果。
检测在卢布林地区输血中心(RBTC)的个体献血检测(IDT)中进行,或在华沙血液学与输血医学研究所(IHTM)对24份样本的混合样本中进行。实时PCR定量检测之前,先采用基于硅胶的方法进行核酸分离(Prepito病毒DNA/RNA、Chemagen和QIAGEN的QIAamp DNA Mini试剂盒)。扩增最初采用自制方法,后来采用商业检测方法(Rotor Gene 6000上的Artus Parvo B19 RG PCR试剂盒)。总共检测了17625份捐献样本:9090份个体样本和8539份混合样本。除了两个实验室都参与的常规外部质量控制项目(荷兰阿姆斯特丹的能力验证研究VQC;苏格兰格拉斯哥的EQA项目)外,还为卢布林的RBTC准备了包含阴性样本、B19V DNA水平非常高的阳性样本以及感染2型基因型的血浆的样本组。
B19V感染频率为1:980份捐献样本,低病毒血症捐献样本最常被检测到(1:1037)。仅鉴定出一份DNA载量可能对血浆制品接受者造成潜在健康风险的捐献样本(1:17625)。在一名捐献者中,B19V DNA被观察到3年零3个月。在感染的急性期或持续期,未观察到临床或实验室症状(外周血形态学、ALT)。由于与病毒基因组多态性相关的病毒载量低估风险,所有B19V检测结果为阳性的捐献样本均不允许临床使用。