Londero Donatella, Fiorino Mauro, Miotti Valeria, de Angelis Vincenzo
Responsible Immunohematology Laboratory, Department of Transfusion Medicine, AOU "S. Maria della Misericordia", Udine, Italy.
Immunohematology. 2011;27(1):25-8.
The known presence of RHD blood group alleles in apparently D– individuals who are positive for C or E antigens leads to an appropriate investigation for the RHD gene on the red blood cells (RBCs) of D– blood donors, thus preventing their RBCs from immunizing D– recipients. Ready-to-use polymerase chain reaction–sequence-specific primer (PCR-SSP) typing kits are available and allow single-sample results. The need to perform this testing on a large number of donors affiliated with the Transfusion Department of Udine (Northern Italy) led to the use of molecular genetic RH blood group typing with PCR-SSP test kits and DNA samples mixed in pools. From a population of 35,000 blood donors screened for D antigen by serologic typing, a total of 235 samples, distributed in pools of 5 DNA samples, were investigated. Positive results were reevaluated by opening the pools and retesting single samples. Validation of DNA-pool typing with commercial kits was done. Among 235 genotyped samples, 12 were found to be PCR positive (5.1%), exhibiting DEL genotype and RHD-CE-D hybrid alleles. Our data demonstrate that the use of a PCR-SSP commercial test kit with pooled samples is a helpful and valid method to correctly detect RHD alleles. As a consequence, we reclassified our donors as carriers of potentially immunogenic alleles.
在C或E抗原呈阳性的看似D抗原阴性个体中，已知存在RHD血型等位基因，这促使对D抗原阴性献血者的红细胞（RBC）进行RHD基因的适当检测，从而防止其红细胞使D抗原阴性受血者产生免疫反应。现成的聚合酶链反应-序列特异性引物（PCR-SSP）分型试剂盒可供使用，并能得出单样本结果。由于需要对乌迪内（意大利北部）输血科的大量献血者进行此项检测，因此采用了分子遗传学RH血型分型方法，使用PCR-SSP检测试剂盒和混合的DNA样本池。在通过血清学分型筛查D抗原的35000名献血者群体中，共调查了235个样本，这些样本以5个DNA样本为一组进行分组。阳性结果通过打开样本池并重新检测单个样本进行重新评估。使用商业试剂盒对DNA样本池分型进行了验证。在235个基因分型样本中，有12个被发现PCR呈阳性（5.1%），表现出DEL基因型和RHD-CE-D杂交等位基因。我们的数据表明，使用带有混合样本的PCR-SSP商业检测试剂盒是正确检测RHD等位基因的一种有用且有效的方法。因此，我们将我们的献血者重新分类为潜在免疫原性等位基因的携带者。
