因妊娠产生抗-D的Rh部分D型基因分析——附3例报告
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摘要
目的研究3例因妊娠产生抗-D Rh部分D型的RHD基因。方法采用间接抗球蛋白试验(IAT)进行Rh D确证试验和抗体鉴定试验,采用序列特异性引物聚合酶链反应(PCR-SSP)技术和PCR直接测序方法对RHD进行基因分型。结果通过血清学试验初步确定3例标本为Rh D变异型,同时均产生了抗-D; PCR-SSP试验检测标本1和标本2为3-6外显子缺失的RHDⅥtypeⅢ型; PCR测序确定标本3为697G>A RHD Va 3型。结论 Rh部分D型在输血或妊娠等免疫刺激后,也存在产生抗体的可能性,以RHDⅥtypeⅢ型多见,在临床输血和新生儿溶血病的预防和诊断过程中需加以注意。
Objective To analyze 3 cases where partial Rh D variant produced anti-D antibody due to pregnancy. Methods Using the indirect antiglobulin test( IAT) to confirm the Rh D phenotype and identify the abnormal antibody. RHD genotyping of these 3 cases was performed by polymerase chain reaction sequence-specific primer( PCR-SSP) and PCR direct sequencing methods. Results By serological tests,3 samples were identified as Rh D variants,and they all produced anti-D antibody. By PCR-SSP test,sample 1 and sample 2 were RHD Ⅵ type Ⅲ,where RHD exon 3—6 were deleted. PCR sequencing results confirmed that sample 3 was RHD Va 3( 697 G>A). Conclusion Partial Rh D variant may produce anti-D antibodies after immunostimulation,such as blood transfusion or pregnancy. This is more common with RHD Ⅵ type Ⅲ. This deserves great attention during the prevention and diagnosis of clinical blood transfusion and hemolytic disease of newborns.
Objective To analyze 3 cases where partial Rh D variant produced anti-D antibody due to pregnancy. Methods Using the indirect antiglobulin test( IAT) to confirm the Rh D phenotype and identify the abnormal antibody. RHD genotyping of these 3 cases was performed by polymerase chain reaction sequence-specific primer( PCR-SSP) and PCR direct sequencing methods. Results By serological tests,3 samples were identified as Rh D variants,and they all produced anti-D antibody. By PCR-SSP test,sample 1 and sample 2 were RHD Ⅵ type Ⅲ,where RHD exon 3—6 were deleted. PCR sequencing results confirmed that sample 3 was RHD Va 3( 697 G>A). Conclusion Partial Rh D variant may produce anti-D antibodies after immunostimulation,such as blood transfusion or pregnancy. This is more common with RHD Ⅵ type Ⅲ. This deserves great attention during the prevention and diagnosis of clinical blood transfusion and hemolytic disease of newborns.
引文
[1]陆松松,陈静娴.Rh(D)变异体的研究进展.临床输血与检验,2010,12(4):374-377.
[2] Avent ND,Reid ME.The Rh blood group system:a review.Blood,2000,95(2):375-387.
[3] Flegel WA,Wangner FF. Molecular genetics of RH. Vox Sang,2000,78(suppl2):109-115.
[4]章旭,李革飞,李剑平.D变异体D IV b型引起重度新生儿溶血病的研究.中华微生物学和免疫学杂志,2014,34(3):224-227.
[5]李丹,邵超鹏,张悦,等.部分D表型同种抗-D 1例及其基因型分析.中国输血杂志,2008,21(9):680-683.
[6]张东民,刘衍春,马玲,等.RHD cat VI type3血型标本的基因分析——附1例报道.中国输血杂志,2012,25(S1):110-110.
[7]尚红,王毓三,申子瑜,等.全国临床检验造作规程.4版.北京:人民卫生出版社,2015:125-126.
[8]杨江存,张崇信,宋耀军,等.聚乙二醇间接抗球蛋白试验临床应用.中国输血杂志,1994,7(4):195-196.
[9] Okuda H,Suganuma H,Tsudo N,et al.Sequence analysis of the spacer region between the RHD and RHCE genes.Biochem Biophys Res Commun,1999,263(2):378-380.
[10] Flegel WA,Wagner FF.Molecular biology of partial D and weak D:implications for blood bank practice. Clin L ab,2002,48(1):53-59.
[11]邵超鹏.RHD研究进展和中国人RHD研究现状分析.中国输血杂志,2003,16(5):354-358.
[12] Esteban R,Montero R,Willy A,et al.The D categoryⅥtype 4allele is prevalent in the Spanish population.Transfusion,2006,46(4):616-623.
[13] Lan JC,Chen Q,Wu DL,et al. Genetic polymorphism of Rh Dnegative associated haplotypes in the Chinese.J Hum Genet,2000,45:224.
[14]左宏莉,何智,罗广平,等.RhDⅥⅢ表型分子生物学研究及其临床意义.中国输血杂志,2008,21:174-176.
[15] Levitt J.Standards for blood banks and transfusion services.29th ed.Bethesda(MD):American Association of Blood Banks,2014.
[16] Daniels G. Rh blood group system.Human blood groups.3rd edition.Oxford:Blackwell,2013:195-274.
[17] Sandler SG,Flegel WA,Westhoff CM,et al.It’s time to phase in RHD genotyping for patients with a serologic weak D phenotype.Transfusion,2015,55:680-689.
[18] Omi T,Takahashi J,Tsudo N,et al.The genomic organization of the partial D category Dva:the presence of a new partial D associated with the Dva phenotype. Biophys Res Commun,1999,254(3):786-794.
[2] Avent ND,Reid ME.The Rh blood group system:a review.Blood,2000,95(2):375-387.
[3] Flegel WA,Wangner FF. Molecular genetics of RH. Vox Sang,2000,78(suppl2):109-115.
[4]章旭,李革飞,李剑平.D变异体D IV b型引起重度新生儿溶血病的研究.中华微生物学和免疫学杂志,2014,34(3):224-227.
[5]李丹,邵超鹏,张悦,等.部分D表型同种抗-D 1例及其基因型分析.中国输血杂志,2008,21(9):680-683.
[6]张东民,刘衍春,马玲,等.RHD cat VI type3血型标本的基因分析——附1例报道.中国输血杂志,2012,25(S1):110-110.
[7]尚红,王毓三,申子瑜,等.全国临床检验造作规程.4版.北京:人民卫生出版社,2015:125-126.
[8]杨江存,张崇信,宋耀军,等.聚乙二醇间接抗球蛋白试验临床应用.中国输血杂志,1994,7(4):195-196.
[9] Okuda H,Suganuma H,Tsudo N,et al.Sequence analysis of the spacer region between the RHD and RHCE genes.Biochem Biophys Res Commun,1999,263(2):378-380.
[10] Flegel WA,Wagner FF.Molecular biology of partial D and weak D:implications for blood bank practice. Clin L ab,2002,48(1):53-59.
[11]邵超鹏.RHD研究进展和中国人RHD研究现状分析.中国输血杂志,2003,16(5):354-358.
[12] Esteban R,Montero R,Willy A,et al.The D categoryⅥtype 4allele is prevalent in the Spanish population.Transfusion,2006,46(4):616-623.
[13] Lan JC,Chen Q,Wu DL,et al. Genetic polymorphism of Rh Dnegative associated haplotypes in the Chinese.J Hum Genet,2000,45:224.
[14]左宏莉,何智,罗广平,等.RhDⅥⅢ表型分子生物学研究及其临床意义.中国输血杂志,2008,21:174-176.
[15] Levitt J.Standards for blood banks and transfusion services.29th ed.Bethesda(MD):American Association of Blood Banks,2014.
[16] Daniels G. Rh blood group system.Human blood groups.3rd edition.Oxford:Blackwell,2013:195-274.
[17] Sandler SG,Flegel WA,Westhoff CM,et al.It’s time to phase in RHD genotyping for patients with a serologic weak D phenotype.Transfusion,2015,55:680-689.
[18] Omi T,Takahashi J,Tsudo N,et al.The genomic organization of the partial D category Dva:the presence of a new partial D associated with the Dva phenotype. Biophys Res Commun,1999,254(3):786-794.