Corcoran A, Kerr S, Elliott G, Koppelman M, Doyle S
Biotrin International, Dublin, Ireland.
Vox Sang. 2007 Oct;93(3):216-22. doi: 10.1111/j.1423-0410.2007.00956.x.
Although parvovirus B19 is a significant blood product contaminant, few methods other than polymerase chain reaction (PCR) have been developed to detect the presence of the virus.
A B19 antigen enzyme immunoassay (EIA) has been developed and the sensitivity of detection is ascertained using dilutions of the B19 capsid protein VP2 and 10-fold dilutions of B19 viraemic serum. Once the assay cut-off was established, a panel of viraemic donations (n = 70) was screened by the antigen EIA. The B19 immunoglobulin M (IgM) and IgG status of these specimens was also determined. During screening of blood donor units by quantitative PCR, 70 individuals were identified with levels of B19 DNA greater than 10(6) IU/ml at the time of blood donation.
The sensitivity of the B19 antigen EIA was estimated to be equivalent to between 10(8) and 10(9) IU/ml B19 DNA or 1-10 pg/ml of recombinant capsid protein. B19 detection was significantly enhanced when viraemic specimens were pretreated with a low pH proprietary reagent. Unlike other virus-detection assays, detection of the B19 antigen was not affected by the presence of B19 IgM or IgG antibodies. In addition, the assay was capable of detecting all three genotypes of human erythrovirus. Combined specimen analysis by the B19 antigen assay and a B19 IgM assay facilitated the detection of 91% of acute B19 infections in the test population.
In combination with B19 IgM detection, application of the B19 antigen EIA is a flexible and efficient method of detecting recent B19 infection and can be used as an alternative to PCR.
尽管细小病毒B19是血液制品的一种重要污染物,但除聚合酶链反应(PCR)外,很少有其他方法可用于检测该病毒的存在。
已开发出一种B19抗原酶免疫测定法(EIA),并使用B19衣壳蛋白VP2的稀释液和B19病毒血症血清的10倍稀释液确定检测灵敏度。一旦确定了检测临界值,就用抗原EIA对一组病毒血症献血样本(n = 70)进行筛查。还测定了这些样本的B19免疫球蛋白M(IgM)和IgG状态。在通过定量PCR对献血单位进行筛查期间,确定有70人在献血时B19 DNA水平高于10(6) IU/ml。
B19抗原EIA的灵敏度估计相当于10(8)至10(9) IU/ml B19 DNA或1 - 10 pg/ml重组衣壳蛋白。当病毒血症样本用低pH专用试剂预处理时,B19检测显著增强。与其他病毒检测方法不同,B19抗原的检测不受B19 IgM或IgG抗体存在的影响。此外,该测定法能够检测人红细胞病毒的所有三种基因型。通过B19抗原测定法和B19 IgM测定法进行联合样本分析,有助于在测试人群中检测出91%的急性B19感染。
与B19 IgM检测相结合,B19抗原EIA的应用是检测近期B19感染的一种灵活高效的方法,可作为PCR的替代方法。