【摘要】：第一部分四川地區(qū)漢族人群Rh(D)變異體分子機(jī)制研究 [目的]研究和分析四川地區(qū)漢族人群Rh(D)變異體的多態(tài)性和分子機(jī)制。為本地區(qū)的Rh(D)變異體研究填補(bǔ)空白,并為Rh血型的準(zhǔn)確鑒定和臨床輸血提供理論基礎(chǔ)和指導(dǎo)。 [方法]首先對所有樣本采用血型血清學(xué)的方法鑒定,初篩陰性的樣本先采用間接抗球蛋白法,以確定弱D或部分D型別樣本,剩余樣本再用吸收放散法,分析Del型別。此外,對樣本的表型進(jìn)行鑒定。用序列特異性引物-聚合酶鏈反應(yīng)方法,對上述樣本進(jìn)行分型確認(rèn)。未能分型成功的疑難樣本進(jìn)行PCR產(chǎn)物直接測序檢測,測序結(jié)果和GenBank上標(biāo)準(zhǔn)序列進(jìn)行比對,以確定變異體的類別。對其雜合性采用PCR-SSP的方法進(jìn)行檢測。 [結(jié)果]血清學(xué)試驗檢出D抗原變異型52例,經(jīng)分子生物學(xué)方法檢測分析,弱D15RHD(G282D)型4例,弱D12RHD(G277E)型1例,弱DRHD(L320L)1例(型別未定),弱DRHD(G263R)1例(型別未定),部分D DVI typeⅢ型2例,DELRHD(K409K)型41例,DELRHD(MlI)型1例,此外發(fā)現(xiàn)新等位基因RHD (A237D) 1例(基因序列號:GU998825)。RHD雜合性檢測結(jié)果顯示1例弱D15和9例DELRHD(K409K)型樣本為RHD+/RHD+純合子,其他均為RHD+/RHD-雜合型。 [結(jié)論]四川地區(qū)漢族人群Rh(D)變異體有豐富的類型和不同分子機(jī)制 第二部分四川地區(qū)漢族人群真正Rh陰性個體基因多態(tài)性研究 [目的]對四川地區(qū)漢族人群RhD陰性個體的基因多態(tài)性進(jìn)行研究. [方法]用PCR-SSP及基因序列測定技術(shù)對經(jīng)間接球蛋白試驗(IAT)、吸收放散試驗檢測均為陰性的樣本進(jìn)行分型。 [結(jié)果]86例RhD陰性的樣本中,包括RHD基因完全缺失個體68例、16例攜帶RHD-CE(2-9)-D等位基因、1例攜帶RHD-CE(8-9)-D等位基因、1例攜帶RHD(711DelC)等位基因。 [結(jié)論]四川地區(qū)漢族人群RhD陰性個體分子機(jī)制具有豐富的多態(tài)性和不同的遺傳背景,其主要為RHD全缺失為主,其次為RHD-CE(2-9)-D型,并存在其他稀有的基因型。
[Abstract]:Part one the molecular mechanism of Rh (D) variant in Sichuan Han population [objective] to study and analyze the polymorphism and molecular mechanism of Rh (D) variant in Sichuan Han population. It provides theoretical basis and guidance for accurate identification of Rh blood group and clinical blood transfusion. [methods] all samples were identified by blood group serological method. Indirect antiglobulin method was used to determine the weak D or part of D type samples, and the remaining samples were analyzed by absorption and elution method. In addition, the phenotype of the sample was identified. The samples were identified by sequence specific primer polymerase chain reaction (PCR). Difficult samples failed to type were detected by direct sequencing of PCR products. The sequencing results were compared with standard sequences on GenBank to determine the type of variants. The heterozygosity was detected by PCR-SSP. [results] 52 cases of D antigen variant were detected by serological test, 4 cases of weak D15RHD (G282D) type and 1 case of weak D12RHD (G277E) type were detected by molecular biological method. There were 1 case of weak DRHD (L320L) (undetermined type), 1 case of weak DRHD (G263R) (undetermined type), 2 cases of partial D DVI type 鈪，

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