Seto Eva, Fernandes Bernard J, Wang Chen, Denomme Gregory A
Department of Pathology & Laboratory Medicine, Mount Sinai Hospital, Canada.
Prenat Diagn. 2007 Nov;27(11):1017-23. doi: 10.1002/pd.1824.
To identify the types of requests, the patterns of testing, and the error rates associated with the genetic identification of fetuses at risk for immune-mediated hemolytic disease.
Retrospective review of the laboratory information system for all fetal blood group genotyping tests performed at Mount Sinai Hospital, Toronto, Canada from January 1997 to December 2006.
Amniotic fluid-derived DNA, from 220 women (243 pregnancies), was tested for one or more antigens (279 tests) when the father was heterozygous for the inferred blood group antigen or was unavailable. The PCR amplification failure rate of amniotic fluid-derived DNA was 5.0%. When the father was considered hemizygous for RHD or was not available, the fetus was positive in 68.6% and 72.7% of cases, respectively. Two amniotic fluid-derived DNA KEL1 SSP-PCR results did not correlate with the result determined from cultured amniocyte DNA.
Paternal RHD hemizygosity may be overestimated. Thus, we recommend that RHD zygosity be established by molecular analysis of the RHD breakpoint. Cultured amniocytes should be reserved for testing if the amniotic fluid-derived DNA is inconclusive due to PCR amplification failure. We use PCR-RFLP genotyping methods for RHCEE/RHCEe (Rh E/e), KEL1/KEL2 (K/k), FYA/FYB (Fy(a/b)), and JKA/JKB (Jk(a/b)).
确定与免疫介导的溶血性疾病风险胎儿的基因鉴定相关的请求类型、检测模式和错误率。
回顾性分析1997年1月至2006年12月在加拿大多伦多西奈山医院进行的所有胎儿血型基因分型检测的实验室信息系统。
当父亲为推断血型抗原的杂合子或无法获得时，对220名女性（243次妊娠）的羊水来源DNA进行了一种或多种抗原检测（279次检测）。羊水来源DNA的PCR扩增失败率为5.0%。当父亲被认为是RHD半合子时或无法获得时，则胎儿分别在68.6%和72.7%的病例中呈阳性。两份羊水来源DNA的KEL1 SSP-PCR结果与培养羊膜细胞DNA测定的结果不相关。
父源性RHD半合子状态可能被高估。因此，我们建议通过RHD断点的分子分析来确定RHD的合子状态。如果由于PCR扩增失败导致羊水来源DNA检测结果不明确，则应保留培养的羊膜细胞用于检测。我们使用PCR-RFLP基因分型方法检测RHCEE/RHCEe（Rh E/e）、KEL1/KEL2（K/k）、FYA/FYB（Fy(a/b)）和JKA/JKB（Jk(a/b)）。
