浙江汉族人群ABO血型等位基因多态性的研究
发布时间：2018-09-06 16:57
【摘要】： 研究背景 ABO血型系统是人类最先发现的重要血型系统,在输血医学和器官移植等方面具有重要意义。经典的血型血清学方法按照抗原的不同可分为A、B、O、AB四种表型,此外ABO血型系统也存在一定的亚型(弱A或弱B)。上世纪九十年代明确了ABO血型基因的核苷酸序列、结构以及一些常见突变点。ABO基因位于染色体9q341-34.2,包含长度大小从28-688bp不等的7个外显子。现一般将A101作为参考序列,B101有7个核苷酸与A101基因不同,导致蛋白上四个氨基酸变化,其中两处替换(L266M和G268A)决定酶的特性。常见的O等位基因上为第6外显子第261位单核苷酸缺失,造成酶活力的丧失。此外也发现少量的261位未缺失的O等位基因。随着分子技术不断完善,现ABO血型基因分型的方法有PCR-RFLP、PCR-SSP、PCR-SSCP、PCR-SSO、PCR-SBT和PCR-GeneScan等,其中最佳的方法是PCR-SBT。到目前为止,通过分子生物学方法已鉴定200多个ABO等位基因。 本研究的目的在于建立ABO血型基因诊断的测序方法(PCRSBT),获取浙江汉族人群ABO血型基因多态性的分布资料,并分析献血人群ABO亚型和不规则抗体分布情况,为临床输血提供更好的技术指导和帮助。实验使用PCR-SBT方法对ABO基因第6、7外显子进行了测序分析,并检测了部分浙江汉族人群标本,发现了三个新的等位基因。 实验方法 1、标本来源 1.1 ABO血型表型分析:2008年7-9月来浙江省血液中心的随机无偿献血者30519名,通过填写征询表确认其民族为汉族。 1.2人群中ABO亚型和不规则抗体调查:2008年来浙江省血液中心的随机无偿献血者,共计标本数为167888人次。 1.3 ABO血型等位基因分布情况:标本来自2008年7-9月浙江省血液中心的随机无偿献血者。所有标本经其知情同意后采集5ml EDTA抗凝血,分别吸取1-2ml血样贮存在-20℃以下备用。标本总数为417例。 2、血清学方法测定 2.1 ABO表型检测:采用PK7200全自动血型检测系统测定ABO血清学表型,参照仪器说明书进行操作。 2.2 ABO亚型鉴定:对于正反定型不一致或者疑是亚型的标本进行鉴定,采用试管法,具体参照中心参比实验有关操作说明进行。 2.3不规则抗体鉴定采用凝集技术在PK7200自动血型检测系统检测。 3、标本DNA提取:采用QIAamp Blood Kit商用抽提试剂盒。 4、ABO基因第6-7外显子测序分析 4.1 PCR扩增:PCR扩增引物序列参照ABO基因序列,采用一对引物对进行5-7外显子扩增,PCR引物序列和扩增条件参照正文,扩增反应体系总体积为25μl。 4.2 PCR产物纯化:每孔中加入核酸外切酶Ⅰ1μl(5U),虾碱性磷酸酶2μl(2U),ABI公司的9700型PCR自动扩增仪进行酶切反应,37℃60min,80℃15min。 4.3测序反应:分别针对外显子6、7设计四个测序引物,以纯化PCR产物为模板按BigDye Sequencing kit(ABI公司)试剂盒操作进行测序反应,ABI公司9700型PCR自动扩增仪扩增,扩增条件为95℃预变性5min,95℃10s,50℃10s,60℃4min,25个循环,冷却至4℃。 4.4测序反应纯化和电泳分析:采用乙醇/醋酸钠法纯化,然后在ABI 3730测序仪上进行测序电泳分析。 4.5测序结果分析和判断:SeqScape V2.5软件进行数据分析。 5磁珠M-270链亲和素方法分离新等位基因的单倍体 当鉴定的新等位基因为杂合子,采用磁珠M-270链亲和素方法(Invitrogen,Wisconsin,USA)分离其单倍体。主要分两步,第一步模板DNA与5′端生物素标记特异性探针反应,生物素探针只与特定的DNA单链结合。第二步将生物素标记单链DNA加入链亲和素化磁珠M-270形成生物素标记DNA-链亲和素包被磁珠,经洗涤、磁选后得到的DNA单链可作为ABO基因外显子6-7的扩增模板。 6、统计学分析:人群基因频率(gene frequency,GF)采用直接计数法计算,即检测出的基因个数/2N(N=标本数)。Hardy-Weinberg(H-W)平衡检验采用x~2检验。p＜0.05表示具有统计学意义。 结果 1、30519例浙江汉族人群ABO血型表型分布情况 采用凝集方法,结果显示A型为9274例,占30.4%。B型为7986例,占26.2%。O型为10542例,占34.5%。AB型为2717例,占8.9%。ABO血型表型分布中O＞A＞B＞AB。 2、ABO亚型与不规则抗体检测情况 从167888例标本中检出8例ABO亚型,ABO亚型频率为4.8/10万。检出43例不规则抗体,不规则抗体检出频率为2.6/万。其中抗M为32例,占74.4%。抗D为4例,占9.3%。冷凝集为3例,占7.0%。抗H、抗C、抗P1、自身抗体各为1例。抗M为最常见的不规则抗体。 3、浙江汉族人群ABO等位基因型分布情况 结果显示外显子6、7序列确定的ABO基因型与表型完全相符。在浙江汉族人群中共发现27种基因型,频率大于1%的为11种。常见的基因型分布为A102/O01为54例(12.95%),A102/O02为34例(8.15%),A102/A102为19例(4.56%);B101/O01为56例(13.43%),B101/O02为38例(9.11%),B101/B101为20例(4.80%),O01/O02为70例(16.78%),O01/O01为44例(10.55%),O02/O02为19例(4.56%),A102/B101为33例(7.91%)、人群中基因型为纯合子的为102例(24.46%),杂合子为315例(75.54%)。 4、浙江汉族人群ABO等位基因分布情况 412例标本中共检测到14个ABO等位基因,包括五个常见的等位基因:A101、A102、B101、O01和O02。此外有6个较少见的等位基因:A205、B110、O04、O05、O07、O50,还有3个新的等位基因:B112、B526C和O743C。频率大于1%的为A101(1.56%)、A102(19.78%)、B101(20.62%)、O01(33.69%)和O02(22.30%)。人群中最常见的等位基因为O01,在A基因中共发现3个等位基因A101、A102、A205,A102频率大于A101,A205发现有2例占0.240%。在B基因中共发现4个等位基因B101,B110、B112、B526C。在O基因中共发现7个等位基因O01、O02、O04、O05、O07、O50、O743C。 5、3个新等位基因情况 在417例标本中发现3个新的等位基因,三个新等位基因的DNA序列已提交GenBank(序列号为FJ599810、FJ638611和FJ851690)。这三个突变点都发生在外显子7区域。 5.1 B112(559T):在一个B/O杂合子个体中发现该等位基因(B112/O01),DNA磁珠纯化技术分离得到单倍体,测序发现在B型单链上存在突变。B112与B101比较在第559位C＞T,导致一个氨基酸改变(Arg187Cys),血清学上未观察到血清学特性的变化。 5.2 B526C:在一个AB个体中发现(A102/B526C),通过磁珠纯化获取单链,结果发现B等位基因在526位G＞C,导致一个氨基酸改变。526位是A和B基因间的差异碱基,在该处A型是C、B型是G。该标本未观察到任何血清学特性变化。 5.3 0743C:在3个标本中发现存在0743C等位基因,其中两个O型杂合子(O01/0743C)和一个B型杂合子(B101/0743C)。与O01序列比较,0743C发生一个突变743G＞C,由于与O01相同存在261位G缺失,在第261位以后发生阅读框架的移动,形成新的终止密码子,提前终止了转移酶的翻译。 结论 1、获取了浙江汉族人群ABO血型表型分布情况。 2、获取了浙江汉族人群ABO亚型与不规则抗体分布情况。 3、首次获取了浙江汉族人群ABO血型等位基因多态性数据。 4、发现了三个新的等位基因。
[Abstract]:Research background
ABO blood group system is an important blood group system first discovered by human beings, which is of great significance in blood transfusion medicine and organ transplantation. Classical blood group serology can be divided into four phenotypes A, B, O and AB according to different antigens. In addition, ABO blood group system also has certain subtypes (weak A or weak B). ABO blood group base was defined in the 1990s. ABO gene is located on chromosome 9q341-34.2 and contains seven exons ranging in length from 28 BP to 688 bp. Al101 is generally used as a reference sequence. There are seven nucleotides in B101 that are different from A101, resulting in changes in four amino acids in the protein. Two substitutions (L266M and G268A) determine enzymes. The most common O allele is the 261 single nucleotide deletion at exon 6, resulting in the loss of enzyme activity. A small number of 261 O alleles are also found. With the development of molecular techniques, ABO genotyping methods are PCR-RFLP, PCR-SSP, PCR-SSCP, PCR-SSO, PCR-SBT and PCR-GeneScan, among which the best are PCR-RFLP, PCR-SSP, PCR-SBT and PCR-GeneScan. The method is PCR-SBT.. Up to now, more than 200 ABO alleles have been identified by molecular biology.
The purpose of this study is to establish a sequencing method for ABO genotyping (PCR-SBT), obtain the distribution of ABO genotype gene polymorphism in Zhejiang Han population, and analyze the distribution of ABO subtypes and irregular antibodies in blood donors, so as to provide better technical guidance and help for clinical blood transfusion. Three new alleles were found in some samples of Zhejiang Han population.
Experimental method
1, specimen sources
1.1 ABO blood group phenotype analysis: 30 519 random blood donors from Zhejiang Blood Center from July to September 2008 were confirmed to be Han nationality by filling in a questionnaire.
1.2 Survey of ABO subtypes and irregular antibodies in the population: A total of 167 888 random blood donors from Zhejiang Blood Center in 2008.
1.3 Distribution of ABO blood group alleles: Specimens were collected from random and unpaid donors in Zhejiang Blood Center from July to September 2008. All specimens were collected with their informed consent and 5 ml EDTA anticoagulant, and 1-2 ml of blood samples were collected and stored below - 20 C for reserve. The total number of specimens was 417.
2. Serological method.
2.1 ABO phenotype detection: PK7200 automatic blood group detection system was used to determine ABO serological phenotype, and the operation was carried out according to the instructions of the instrument.
2.2 ABO subtype identification: For samples with inconsistent positive and negative stereotypes or suspected subtypes, test tube method was used, with specific reference to the operation instructions of the central reference experiment.
2.3 the identification of irregular antibodies was detected by agglutination technique in PK7200 automatic blood typing system.
3, specimen DNA extraction: QIAamp Blood Kit commercial extraction kit.
4, sequencing analysis of exon 6-7 of ABO gene.
4.1 PCR amplification: PCR amplification primer sequence refers to ABO gene sequence, using a pair of primers for 5-7 exon amplification, PCR primer sequence and amplification conditions refer to the text, the total volume of the amplification reaction system is 25 mu L.
4.2 PURIFICATION OF PCR PRODUCTS: Each pore was digested with exonuclease_1_ (5U), shrimp alkaline phosphatase 2_ (2U) and ABI company's 9700 PCR automatic amplifier, at 37 60 min and 80 15 min.
4.3 Sequencing reaction: Four primers were designed for exon 6 and exon 7 respectively. The purified PCR products were used as templates for sequencing according to BigDye Sequencing kit (ABI Company) kit. The amplification conditions were pre-denatured at 95 C for 5 minutes, pre-denatured at 95 C for 10 seconds, 10 seconds at 50 C for 4 minutes, and cooled to 4 C for 25 cycles.
4.4 Sequencing reaction purification and electrophoresis analysis: Purification by ethanol/sodium acetate method, and then sequencing electrophoresis analysis on ABI 3730 sequencer.
4.5 analysis and judgement of sequencing results: SeqScape V2.5 software for data analysis.
5 magnetic beads M-270 chain avidin method for isolation of haploid from new alleles
When a new allele was identified as a heterozygote, the haploid was isolated by magnetic bead M-270 chain avidin method (Invitrogen, Wisconsin, USA). The first step was to react the template DNA with a 5'-terminal biotin-labeled specific probe. The biotin probe was only bound to a specific DNA single strand. The second step was to add the biotin-labeled single strand DNA to the chain affinity. The biotin-labeled DNA-chain avidin coated magnetic beads were formed by the biotin-labeled magnetic beads M-270. After washing and magnetic separation, the DNA single strands could be used as amplification templates for exon 6-7 of ABO gene.
6. Statistical analysis: The population gene frequency (GF) was calculated by direct counting method, i.e. the number of detected genes / 2N (N = the number of samples). Hardy-Weinberg (H-W) balance test was performed by X-2 test. P < 0.05 showed statistical significance.
Result
Distribution of ABO blood group phenotypes in 130519 Han population in Zhejiang
Agglutination was used in 9274 cases (30.4%) of type A, 7986 cases (26.2%) of type B, 10542 cases (34.5%) of type O, 2717 cases (8.9%) of type ABO, and O > A > B > AB.
2, the detection of ABO subtypes and irregular antibodies.
ABO subtypes were detected in 8 out of 16788 specimens. The frequency of ABO subtypes was 48/100 000. 43 irregular antibodies were detected. The frequency of irregular antibodies was 26/10 000. Among them, 32 were anti-M, accounting for 74.4%. Anti-D was 4, accounting for 9.3%. Condensation was 3, accounting for 7.0%. Anti-H, anti-C, anti-P1, and autoantibodies were the most common irregular antibodies.
3, the distribution of ABO alleles in Zhejiang Han population.
The results showed that the ABO genotypes determined by exon 6 and exon 7 were completely consistent with the phenotype. Twenty-seven genotypes were found in Zhejiang Han population with frequencies greater than 1%. The common genotypes were A102/O01 in 54 cases (12.95%), A102/O02 in 34 cases (8.15%), A102/A102 in 19 cases (4.56%), B101/O01 in 56 cases (13.43%), B101/O02 in 38 cases (9.11%) and B101/O02 in 38 cases (9.11%). 01/B101 was 20 cases (4.80%), O01/O02 was 70 cases (16.78%), O01/O01 was 44 cases (10.55%), O02/O02 was 19 cases (4.56%), A102/B101 was 33 cases (7.91%), 102 cases (24.46%) were homozygotes and 315 cases (75.54%) were heterozygotes.
4, the distribution of ABO alleles in Zhejiang Han population.
Fourteen ABO alleles were detected in 412 specimens, including five common alleles: A101, A102, B101, O01 and O02. In addition, six rare alleles were found: A205, B110, O04, O05, O07, O50, and three new alleles: B112, B526C and O743C. Frequencies greater than 1% were A101 (1.56%), A102 (19.78%), B101 (20.62%), O01 (33.69%) and O02 (22.3%). The most common allele in the population was O01. Three alleles A101, A102, A205, A102 were found in A gene with frequencies higher than A101, and 2 alleles in A205 accounted for 0.240%. Four alleles B101, B110, B112 and B526C were found in B gene. Seven alleles O01, O02, O04, O05, O07, O50, O743C.
5,3 new alleles
Three new alleles were found in 417 specimens. The DNA sequences of three new alleles were submitted to GenBank (FJ599810, FJ638611 and FJ851690). All three mutations occurred in exon 7 region.
5.1 B112 (559T): The allele (B112/O01) was found in a B/O heterozygote, and the haploid was isolated by magnetic bead purification technique. Mutations were found in the B-type single strand. B112 was compared with B101 at position 559 C > T, resulting in an amino acid change (Arg187Cys). Serological changes were not observed.
5.2 B526C: A single strand was found in an AB individual (A102/B526C) and purified by magnetic beads. It was found that the B allele at position 526 G > C resulted in an amino acid change.
5.3 0743C: Two O-type heterozygotes (O01/0743C) and one B-type heterozygote (B101/0743C) were found in three specimens. Compared with O01 sequence, a mutation 743G > C occurred in 0743C. Because of 261 G deletion, the reading frame shifted after position 261, forming a new termination codon and terminating ahead of time. The translation of the transferase was stopped.
conclusion
1, the distribution of ABO blood group phenotypes in Zhejiang Han population was obtained.
2, we obtained the distribution of ABO subtypes and irregular antibodies in Zhejiang Han population.
3, we obtained the ABO blood group allele polymorphism data of Han nationality in Zhejiang for the first time.
4, three new alleles were found.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R394
本文编号：2226973
[Abstract]:Research background
ABO blood group system is an important blood group system first discovered by human beings, which is of great significance in blood transfusion medicine and organ transplantation. Classical blood group serology can be divided into four phenotypes A, B, O and AB according to different antigens. In addition, ABO blood group system also has certain subtypes (weak A or weak B). ABO blood group base was defined in the 1990s. ABO gene is located on chromosome 9q341-34.2 and contains seven exons ranging in length from 28 BP to 688 bp. Al101 is generally used as a reference sequence. There are seven nucleotides in B101 that are different from A101, resulting in changes in four amino acids in the protein. Two substitutions (L266M and G268A) determine enzymes. The most common O allele is the 261 single nucleotide deletion at exon 6, resulting in the loss of enzyme activity. A small number of 261 O alleles are also found. With the development of molecular techniques, ABO genotyping methods are PCR-RFLP, PCR-SSP, PCR-SSCP, PCR-SSO, PCR-SBT and PCR-GeneScan, among which the best are PCR-RFLP, PCR-SSP, PCR-SBT and PCR-GeneScan. The method is PCR-SBT.. Up to now, more than 200 ABO alleles have been identified by molecular biology.
The purpose of this study is to establish a sequencing method for ABO genotyping (PCR-SBT), obtain the distribution of ABO genotype gene polymorphism in Zhejiang Han population, and analyze the distribution of ABO subtypes and irregular antibodies in blood donors, so as to provide better technical guidance and help for clinical blood transfusion. Three new alleles were found in some samples of Zhejiang Han population.
Experimental method
1, specimen sources
1.1 ABO blood group phenotype analysis: 30 519 random blood donors from Zhejiang Blood Center from July to September 2008 were confirmed to be Han nationality by filling in a questionnaire.
1.2 Survey of ABO subtypes and irregular antibodies in the population: A total of 167 888 random blood donors from Zhejiang Blood Center in 2008.
1.3 Distribution of ABO blood group alleles: Specimens were collected from random and unpaid donors in Zhejiang Blood Center from July to September 2008. All specimens were collected with their informed consent and 5 ml EDTA anticoagulant, and 1-2 ml of blood samples were collected and stored below - 20 C for reserve. The total number of specimens was 417.
2. Serological method.
2.1 ABO phenotype detection: PK7200 automatic blood group detection system was used to determine ABO serological phenotype, and the operation was carried out according to the instructions of the instrument.
2.2 ABO subtype identification: For samples with inconsistent positive and negative stereotypes or suspected subtypes, test tube method was used, with specific reference to the operation instructions of the central reference experiment.
2.3 the identification of irregular antibodies was detected by agglutination technique in PK7200 automatic blood typing system.
3, specimen DNA extraction: QIAamp Blood Kit commercial extraction kit.
4, sequencing analysis of exon 6-7 of ABO gene.
4.1 PCR amplification: PCR amplification primer sequence refers to ABO gene sequence, using a pair of primers for 5-7 exon amplification, PCR primer sequence and amplification conditions refer to the text, the total volume of the amplification reaction system is 25 mu L.
4.2 PURIFICATION OF PCR PRODUCTS: Each pore was digested with exonuclease_1_ (5U), shrimp alkaline phosphatase 2_ (2U) and ABI company's 9700 PCR automatic amplifier, at 37 60 min and 80 15 min.
4.3 Sequencing reaction: Four primers were designed for exon 6 and exon 7 respectively. The purified PCR products were used as templates for sequencing according to BigDye Sequencing kit (ABI Company) kit. The amplification conditions were pre-denatured at 95 C for 5 minutes, pre-denatured at 95 C for 10 seconds, 10 seconds at 50 C for 4 minutes, and cooled to 4 C for 25 cycles.
4.4 Sequencing reaction purification and electrophoresis analysis: Purification by ethanol/sodium acetate method, and then sequencing electrophoresis analysis on ABI 3730 sequencer.
4.5 analysis and judgement of sequencing results: SeqScape V2.5 software for data analysis.
5 magnetic beads M-270 chain avidin method for isolation of haploid from new alleles
When a new allele was identified as a heterozygote, the haploid was isolated by magnetic bead M-270 chain avidin method (Invitrogen, Wisconsin, USA). The first step was to react the template DNA with a 5'-terminal biotin-labeled specific probe. The biotin probe was only bound to a specific DNA single strand. The second step was to add the biotin-labeled single strand DNA to the chain affinity. The biotin-labeled DNA-chain avidin coated magnetic beads were formed by the biotin-labeled magnetic beads M-270. After washing and magnetic separation, the DNA single strands could be used as amplification templates for exon 6-7 of ABO gene.
6. Statistical analysis: The population gene frequency (GF) was calculated by direct counting method, i.e. the number of detected genes / 2N (N = the number of samples). Hardy-Weinberg (H-W) balance test was performed by X-2 test. P < 0.05 showed statistical significance.
Result
Distribution of ABO blood group phenotypes in 130519 Han population in Zhejiang
Agglutination was used in 9274 cases (30.4%) of type A, 7986 cases (26.2%) of type B, 10542 cases (34.5%) of type O, 2717 cases (8.9%) of type ABO, and O > A > B > AB.
2, the detection of ABO subtypes and irregular antibodies.
ABO subtypes were detected in 8 out of 16788 specimens. The frequency of ABO subtypes was 48/100 000. 43 irregular antibodies were detected. The frequency of irregular antibodies was 26/10 000. Among them, 32 were anti-M, accounting for 74.4%. Anti-D was 4, accounting for 9.3%. Condensation was 3, accounting for 7.0%. Anti-H, anti-C, anti-P1, and autoantibodies were the most common irregular antibodies.
3, the distribution of ABO alleles in Zhejiang Han population.
The results showed that the ABO genotypes determined by exon 6 and exon 7 were completely consistent with the phenotype. Twenty-seven genotypes were found in Zhejiang Han population with frequencies greater than 1%. The common genotypes were A102/O01 in 54 cases (12.95%), A102/O02 in 34 cases (8.15%), A102/A102 in 19 cases (4.56%), B101/O01 in 56 cases (13.43%), B101/O02 in 38 cases (9.11%) and B101/O02 in 38 cases (9.11%). 01/B101 was 20 cases (4.80%), O01/O02 was 70 cases (16.78%), O01/O01 was 44 cases (10.55%), O02/O02 was 19 cases (4.56%), A102/B101 was 33 cases (7.91%), 102 cases (24.46%) were homozygotes and 315 cases (75.54%) were heterozygotes.
4, the distribution of ABO alleles in Zhejiang Han population.
Fourteen ABO alleles were detected in 412 specimens, including five common alleles: A101, A102, B101, O01 and O02. In addition, six rare alleles were found: A205, B110, O04, O05, O07, O50, and three new alleles: B112, B526C and O743C. Frequencies greater than 1% were A101 (1.56%), A102 (19.78%), B101 (20.62%), O01 (33.69%) and O02 (22.3%). The most common allele in the population was O01. Three alleles A101, A102, A205, A102 were found in A gene with frequencies higher than A101, and 2 alleles in A205 accounted for 0.240%. Four alleles B101, B110, B112 and B526C were found in B gene. Seven alleles O01, O02, O04, O05, O07, O50, O743C.
5,3 new alleles
Three new alleles were found in 417 specimens. The DNA sequences of three new alleles were submitted to GenBank (FJ599810, FJ638611 and FJ851690). All three mutations occurred in exon 7 region.
5.1 B112 (559T): The allele (B112/O01) was found in a B/O heterozygote, and the haploid was isolated by magnetic bead purification technique. Mutations were found in the B-type single strand. B112 was compared with B101 at position 559 C > T, resulting in an amino acid change (Arg187Cys). Serological changes were not observed.
5.2 B526C: A single strand was found in an AB individual (A102/B526C) and purified by magnetic beads. It was found that the B allele at position 526 G > C resulted in an amino acid change.
5.3 0743C: Two O-type heterozygotes (O01/0743C) and one B-type heterozygote (B101/0743C) were found in three specimens. Compared with O01 sequence, a mutation 743G > C occurred in 0743C. Because of 261 G deletion, the reading frame shifted after position 261, forming a new termination codon and terminating ahead of time. The translation of the transferase was stopped.
conclusion
1, the distribution of ABO blood group phenotypes in Zhejiang Han population was obtained.
2, we obtained the distribution of ABO subtypes and irregular antibodies in Zhejiang Han population.
3, we obtained the ABO blood group allele polymorphism data of Han nationality in Zhejiang for the first time.
4, three new alleles were found.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R394
【引证文献】
相关期刊论文 前1条
1 冯君;应晓君;;浙江汉族人ABO血型与肺癌相关性分析[J];哈尔滨医药;2013年02期
，本文编号：2226973
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2226973.html
最近更新
教材专著
