黑龍江地區(qū)人群HLA-A、B、DRB1高分辨等位基因及單體型多態(tài)性研究①

李　鑫　丁　鐫　王　鑫　侯　玲　劉　穎　劉　杰

(哈爾濱市血液中心，哈爾濱150056)

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黑龍江地區(qū)人群HLA-A、B、DRB1高分辨等位基因及單體型多態(tài)性研究①

李鑫丁鐫王鑫侯玲劉穎②劉杰②

(哈爾濱市血液中心，哈爾濱150056)

[摘要]目的：研究中國(guó)造血干細(xì)胞捐獻(xiàn)者資料庫(kù)黑龍江分庫(kù)無(wú)關(guān)捐獻(xiàn)者HLA-A、B、DRB1高分辨等位基因和單體型頻率分布特征。方法：采用PCR-測(cè)序分型技術(shù)(SBT)對(duì)13 670名隨機(jī)、無(wú)血緣關(guān)系、健康的中華骨髓庫(kù)黑龍江分庫(kù)造血干細(xì)胞捐獻(xiàn)者進(jìn)行HLA-A、B、DRB1高分辨基因分型，利用計(jì)數(shù)法和最大似然性原理方法分別計(jì)算HLA-A、B、DRB1等位基因及單體型頻率。結(jié)果：共觀察到286個(gè)HLA等位基因，其中頻率>0.1的A、B、DRB1座位特異性分別為A* 02∶01、A*24∶02、A*11∶01、DR*07∶01、DR*09∶01、DR*15∶01。1 087條A-B單體型中，22條單體型的頻率高于0.01,267條單體型呈現(xiàn)顯著的正連鎖不平衡(ALD>0,HF≥1.09×10-4,χ2>3.84)，12條表現(xiàn)為強(qiáng)連鎖不平衡(RLD>0.80)；1 329條B-DRB1單體型中，19條單體型的頻率高于0.01,357條單體型呈現(xiàn)顯著的正連鎖不平衡，18條表現(xiàn)為強(qiáng)連鎖不平衡(RLD>0.80)；4 428條A-B-DRB1單體型中，1 348條單體型有意義(頻率≥1.09×10-4)，17條單體型頻率高于0.005。結(jié)論：獲得黑龍江地區(qū)人群HLA-A、B、DRB1高分辨等位基因頻率和單體型分布數(shù)據(jù)及相關(guān)遺傳參數(shù)，等位基因和單體型的分布特征接近北方漢族人群，但具有其自身分布特征。

HLA系統(tǒng)是迄今所知人類基因中多態(tài)性最復(fù)雜的遺傳系統(tǒng)，其等位基因頻率及其連鎖不平衡特點(diǎn)在不同種族、民族、地域存在明顯差異。其多態(tài)性及分布差異是自然選擇造成的，它保證了種群能產(chǎn)生對(duì)各種病原體特異的免疫應(yīng)答，以維持群體的穩(wěn)定性。因此，一個(gè)群體高分辨水平的HLA多態(tài)性的研究對(duì)組織器官移植尋找合適供者、群體遺傳學(xué)、人類學(xué)和法醫(yī)學(xué)至關(guān)重要。目前有關(guān)用骨髓庫(kù)樣本檢測(cè)HLA基因多態(tài)性在中國(guó)人群中的報(bào)道盡管已有很多，但大部分或是小樣本數(shù)據(jù)調(diào)查，或是低分辨分型數(shù)據(jù)的研究。我們采用SBT分型技術(shù)對(duì)2009年至2014年13 670例中國(guó)干細(xì)胞庫(kù)黑龍江地區(qū)自愿捐獻(xiàn)者進(jìn)行HLA-A、B、DRB1高分辨分型，得到了較為可靠、完整的大樣本HLA群體分型數(shù)據(jù)，統(tǒng)計(jì)分析了HLA等位基因頻率、單體型頻率以及連鎖不平衡結(jié)果等相關(guān)遺傳參數(shù)資料，現(xiàn)報(bào)道如下。

1材料與方法

1.1材料

1.1.1標(biāo)本來(lái)源13 670份標(biāo)本全部來(lái)自2009年至2014年黑龍江地區(qū)健康、無(wú)血緣關(guān)系、18~55周歲造血干細(xì)胞捐獻(xiàn)者。

1.1.2主要試劑和儀器TIANamp Blood DNA Kit血液基因組DNA提取試劑(美國(guó) Tiangen公司)，PCR-SBT 基因分型試劑盒(德國(guó)ROSE公司)，9 700 PCR 擴(kuò)增儀，Bio-Rad PCR 擴(kuò)增儀，3730xl DNA Analyzer測(cè)序儀。

1.2實(shí)驗(yàn)方法外周靜脈血樣(EDTA抗凝)5 ml,應(yīng)用TIANamp Blood DNA Kit血液基因組DNA提取試劑(美國(guó) Tiangen公司)提取基因組DNA，檢測(cè)DNA濃度在40~100 ng/μl，OD260/280比值為1.6~1.8。采用PCR-SBT方法對(duì)HLA-A、B、DRB1 進(jìn)行常規(guī)測(cè)序分型，操作步驟嚴(yán)格按試劑說明書進(jìn)行。其中對(duì)屬于HLAⅠ類基因的A、B位點(diǎn)進(jìn)行第2、3和4外顯子雙向測(cè)序，而對(duì)于HLAⅡ類基因的DRB1進(jìn)行第2外顯子雙向測(cè)序.對(duì)無(wú)法確定高分辨分型結(jié)果的樣本，則選擇序列特異性測(cè)序引物(Sequence specific sequencing primer,SSSP)對(duì)該樣本再次測(cè)序，獲得相應(yīng)的高分辨結(jié)果。測(cè)序反應(yīng)純化后的產(chǎn)物置于3730xl DNA Analyzer測(cè)序儀上進(jìn)行電泳分析，導(dǎo)出的結(jié)果采用序列分析軟件ATF-loader進(jìn)行分析。

1.3統(tǒng)計(jì)學(xué)處理

1.3.1等位基因頻率和單體型頻率采用直接計(jì)數(shù)法和最大數(shù)學(xué)預(yù)期值算法(expectation-maximization,EM)通過Arlequin3.1軟件分別計(jì)算等位基因頻率和單體型頻率，同時(shí)對(duì)各基因位點(diǎn)作Hardy-Weinberg平衡檢驗(yàn)[1]。

1.3.2最小群體數(shù)量的估計(jì)在由n個(gè)個(gè)體組成的群體中，某單體型只出現(xiàn)一次，該單體型頻率HP=1/2n，但并非所有頻率≥1/2n的單體型都是可靠的。由此在隨機(jī)人群中，如果要求有P的把握在群體中檢測(cè)到該單體型，所需要的群體最少數(shù)量為N，n= log(1-P)/log(1-2 hP+HP2)。如果把P設(shè)置在95%區(qū)間，則n=-1.3010/log(1-2 hP+HP2)[2]。

1.3.3連鎖不平衡參數(shù)計(jì)算兩座位HLA單體型連鎖不平衡參數(shù)的計(jì)算采用標(biāo)準(zhǔn)表格法，包括絕對(duì)△值(ALD)、最大△值(mlD)和相對(duì)△值(RLD)，計(jì)算公式：△ij=fij-fi×fj;△max=fij(△ij<0),△max=fi×(1-fj)(△ij≥0且fi

1.3.4頻率分布的熵值(Entropy)計(jì)算在信息學(xué)理論中，頻率分布的熵值是對(duì)包括多個(gè)子項(xiàng)的項(xiàng)目所含信息量的度量，具體到HLA領(lǐng)域，是對(duì)1個(gè)或1組緊密連鎖的HLA基因座位多態(tài)性高低的度量[3]，計(jì)算公式：E=-∑fi×log2fi(fi為HLA基因或單體型的頻率)。

2結(jié)果

2.1HLA-A、B、DRB1等位基因頻率13670份標(biāo)本共檢測(cè)到HLA-A、B、DRB1基因座位上286種HLA編碼基因(見表1)。HLA-A、B、DRB1基因座分別檢出等位基因80、136和70個(gè)基因。A、B、DRB1基因座HW吻合度的檢驗(yàn)結(jié)果均為P>0.05，表明本研究來(lái)源群體的基因分布符合Hardy-Weinberg平衡定律。各座位上等位基因頻率的分布情況顯示，E(B)>E(DRB1)>E(A)，表明HLA基因座位B、DRB1、A的多態(tài)性程度依次降低(見表2)。

2.2最小群體數(shù)量的估計(jì)在調(diào)查的群體樣本中，有95%可信度的單體型頻率≥1.09×10-4認(rèn)為是可靠的，小于此數(shù)值的單體型頻率，則需要在更大樣本的群體資料中進(jìn)行估計(jì)。

2.3HLA單體型頻率分布情況見表3，觀察到1087條HLA-A-B、1329條HLA-B-DRB1和4428條HLA-A-B-DRB1單體型，分別占單體型理論總數(shù)的9.99%(1 087/10 880)、13.96%(1 329/9 520)、0.58%(4 428/761 600)。4428條A-B-DRB1單體型中“可靠”的(頻率≥1.09×10-4)有1 348條，占單體型總數(shù)的30.44%(1 348/4 428)，但累計(jì)頻率高達(dá)85.17%，其中頻率>0.001有158條，頻率最高的10條HLA-A-B、B-DRB1和A-B-DRB1單體型及其頻率數(shù)值見表4。

2.4連鎖不平衡參數(shù)本組資料僅選取頻率≥1.09×10-4的有意義單體型計(jì)算連鎖不平衡參數(shù)。結(jié)果顯示，分別有267條HLA-A-B單體型和357條HLA-B-DRB1單體型呈現(xiàn)顯著的正連鎖不平衡(ALD>0,HF≥1.09×10-4,χ2>3.84),其中A*02∶53N-B*27∶04、A*25∶01-B*18∶01等12條A-B單體型和B*42∶01-DR*13∶02、B*27∶24-DR*04∶05等18條B-DRB1單體型表現(xiàn)為強(qiáng)連鎖不平衡(RLD>0.80)。見表5。表1黑龍江地區(qū)HLA-A、B、DRB1高分辨等位基因頻率

Tab.1High resolution allele frequencies of HLA-A,B,DRB1 in population from Heilongjiang region

HLA-AGeneFrequecyHLA-BGeneFrequecyHLA-BGeneFrequecyHLA-DRB1GeneFrequecy02∶010.16550813∶020.09513527∶250.00021907∶010.14239224∶020.15032946∶010.07092251∶070.00021909∶010.14074611∶010.14312440∶010.05746253∶010.00021915∶010.13668630∶010.08953915∶010.05665756∶030.00021912∶020.07849333∶030.07867651∶010.05665739∶090.00018311∶010.05449902∶060.07183640∶060.05193944∶050.00018308∶030.05193902∶070.05775413∶010.04758655∶120.00014613∶020.04319701∶010.04890358∶010.04718415∶1980.00011003∶010.03939331∶010.04151444∶030.04019815∶680.00011004∶050.03924703∶010.03628452∶010.03675935∶040.00011015∶020.0339826∶010.02673748∶010.03511340∶110.00011014∶050.02717632∶010.01784954∶010.03120042∶020.00011012∶01∶01G0.02633502∶030.01554515∶110.02904251∶060.00011001∶010.01964268∶010.00716935∶01∶01G0.02699307∶100.00007304∶060.01938602∶050.00669340∶020.02509115∶100.00007313∶010.01806929∶010.0066207∶020.02275115∶190.00007316∶020.01572811∶020.00621815∶180.02110539∶310.00007310∶010.01547202∶100.00508457∶010.01799640∶780.00007304∶010.01269203∶020.00292615∶020.01682544∶270.00007304∶030.01225333∶010.00248738∶020.01667951∶220.00007314∶01∶01G0.01152223∶010.00234155∶020.01620351∶360.00007308∶020.00771824∶200.00226837∶010.01525273∶010.00007314∶030.00720624∶080.001539∶010.01422807∶480.00003711∶040.00636430∶040.00131735∶030.01393613∶160.00003714∶040.00625569∶010.00106167∶010.01075315∶090.00003704∶040.00592566∶010.00095150∶010.01027815∶150.00003701∶020.00303668∶020.00087851∶020.01013215∶1520.00003714∶070.00274324∶070.00076808∶010.00991215∶1780.00003704∶100.00259729∶020.00076815∶270.00888815∶340.00003704∶070.00234102∶53N0.00069544∶02∶01G0.00735215∶660.00003704∶080.00201224∶040.00058527∶040.00596215∶860.00003704∶020.00186511∶030.00054938∶010.00585227∶360.00003713∶120.00179226∶030.00051227∶050.00566927∶400.00003711∶060.0013930∶020.00043907∶05∶01G0.00563335∶110.00003715∶040.0013924∶030.00040214∶020.00398735∶120.00003708∶010.00102402∶480.00036615∶070.00369435∶140.00003714∶120.00087824∶100.00036640∶030.00354837∶020.00003716∶010.00084102∶090.00032918∶010.00292639∶040.00003708∶040.00080526∶020.00021915∶170.00281639∶060.00003713∶070.00076834∶010.00021956∶010.00245139∶240.00003713∶030.000695

(轉(zhuǎn)下頁(yè))

(續(xù)表1)

HLA-AGeneFrequecyHLA-BGeneFrequecyHLA-BGeneFrequecyHLA-DRB1GeneFrequecy02∶110.00018335∶080.00230440∶1250.00003708∶090.00062225∶010.00018349∶010.00215840∶250.00003711∶030.00043902∶020.00014615∶050.00212140∶430.00003714∶020.00040202∶420.00014635∶020.00201240∶480.00003714∶180.00032968∶240.00014627∶070.00190240∶550.00003715∶060.00021902∶900.00011015∶250.00182940∶590.00003714∶060.00018324∶300.00011041∶010.00153640∶840.00003711∶280.00011030∶180.00011039∶050.00135346∶160.00003713∶500.00011074∶01∶01G0.00011045∶010.0012851∶040.00003715∶030.00011002∶170.00007315∶320.00124451∶090.00003703∶270.00007303∶080.00007359∶010.00106152∶110.00003711∶190.00007311∶060.00007348∶030.00102454∶160.00003713∶050.00007323∶260.00007355∶010.00102454∶170.00003714∶100.00007333∶080.00007314∶010.00095156∶150.00003714∶250.00007368∶380.00007315∶580.00084157∶020.00003714∶490.00007374∶030.00007340∶400.00080557∶030.00003703∶350.00003792∶890.00007355∶040.00080554∶160.00003704∶130.00003701∶030.00003735∶050.00076854∶170.00003711∶200.00003702∶040.00003751∶080.00073256∶150.00003711∶570.00003702∶1450.00003715∶460.00065857∶020.00003712∶030.00003702∶200.00003742∶010.00054957∶030.00003712∶050.00003702∶360.00003781∶01∶01G0.00054981∶010.00003712∶160.00003702∶930.00003715∶130.00051212∶200.00003711∶040.00003747∶010.00043913∶060.00003711∶190.00003755∶070.00043913∶130.00003711∶360.00003727∶020.00040213∶190.00003711∶390.00003781∶020.00040214∶220.00003711∶880.00003715∶120.00036614∶290.00003724∶1210.00003715∶350.00036615∶050.00003724∶170.00003715∶080.00032915∶310.00003724∶210.00003727∶240.00032915∶050.00003724∶280.00003715∶030.00029315∶310.00003724∶320.00003715∶290.00025624∶680.00003727∶060.00025626∶080.00003741∶020.00025626∶180.00003756∶040.00025626∶200.00003715∶210.00021929∶100.00003715∶380.00021932∶030.00003718∶020.00021974∶020.00003727∶030.000219

Note：HLA-A*74∶01∶01G includes 74∶01,74∶02；HLA-B*07∶05∶01G includes 07∶05，07∶06；HLA-B*35∶01∶01G includes 35∶01，35∶42；HLA-B*81∶01∶01G includes 81∶01，81∶02；HLA-B*44∶02∶01G includes 44∶02，44∶19N；HLA-DRB1*12∶01∶01G includes 12∶01，12∶06，12∶10，12∶17；HLA-DRB1*14∶01∶01G includes 14∶01，14∶54.

表2黑龍江地區(qū)HLA-A、B、DRB1基因頻率分布

Tab.2Frequency distribution for the alleles of HLA-A，B，DRB1

Frequencyrange(×10-2)HLA-AnCumfre(%)HLA-BnCumfre(%)HLA-DRnCumfre(%)>10345.900341.981-101048.462789.411751.90.1-1124.57268.95155.37<0.1551.07831.64350.75Total8013670100EntropyE=3.74E=5.05E=4.19

表3黑龍江地區(qū)HLA-A-B、B-DRB1、A-B-DRB1單體型頻率分布

Tab.3Haplotype frequency distribution of HLA-A,B,DRB1 in population from Heilongjiang region

Frequencyrange(×10-4)HLA-A-BnCumfre(%)HLA-B-DRB1nCumfre(%)HLA-A-B-DRB1nCumfre(%)100-10002241.161938.38614.4410-10013542.4714640.3715235.6<1093016.37116421.25427049.96Notobserved97930819107571720Total108809520761600EntropyE=7.55E=7.84E=9.86

表5高度連鎖不平衡的HLA-A-B、HLA-B-DRB1單體型

Tab.5Positive linkage disequilibrium of HLA-A-B，B-DRB1 haplotypes

ABDRHF(×10-6)ALDmlDRLDχ202∶53N27∶040.0001100.0000940.0000941.0018.0125∶0118∶010.0001100.0001000.0001001.0030.5833∶0114∶020.0024510.0024410.0024770.9916537.0730∶0113∶020.0789720.0704540.0810210.8719337.8629∶0245∶010.0006580.0006340.0007440.85473.8230∶0414∶010.0008050.0008040.0009500.8514133.7429∶0107∶05∶01G0.0047170.0046800.0055960.8416254.6102∶4851∶010.0001830.0001830.0002190.8411419.5502∶4867∶010.0001830.0001830.0002190.8411419.5568∶0227∶030.0001830.0001830.0002190.844756.9202∶0550∶010.0055910.0055220.0066240.8312328.1534∶0115∶210.0001830.0001740.0002100.8398.2142∶0113∶020.0005490.0005250.0005251.00332.6427∶2404∶050.0003290.0003160.0003161.00220.2615∶2115∶020.0002190.0002190.0002191.0013635.8635∶0308∶010.0001100.0000950.0000951.0018.3640∶0208∶040.0001100.0001100.0001101.006848.3254∶0110∶010.0001100.0001070.0001071.0093.3915∶1312∶020.0005010.0004610.0004720.98156.8415∶3215∶040.0010970.0010950.0012420.8819017.4014∶0201∶020.002670.0026580.0030240.8816068.7437∶0108∶020.0001610.0001580.0001800.88249.0235∶0211∶040.0017550.0017420.0019990.876535.5415∶4615∶010.0005740.0004840.0005680.8582.5615∶1713∶020.0024060.0022840.0026940.851229.2681∶01∶01G15∶010.0004750.0004000.0004740.8467.5515∶2915∶010.0002190.0001840.0002210.8330.6508∶0103∶010.0082870.0078970.0095220.834590.6315∶3811∶010.0001830.0001710.0002070.8370.9113∶0207∶010.0808780.0673320.0815890.8311790.63

表4黑龍江地區(qū)10條最常見HLA-A-B、B-DRB1、A-B-DRB1單體型頻率(×10-6)

Tab.4Major 10 HLA-A-B,B-DRB1,A-B-DRB1 haplotypes and their haplotype frequencies(×10-6)

HLA-A-BHFB-DRB1HFA-B-DRB1HFA*30∶01-B*13∶0278972B*13∶02-DR*07∶0180878A*30∶01-B*13∶02-DR*07∶0167302A*02∶07-B*46∶0140870B*46∶01-DR*09∶0134388A*02∶07-B*46∶01-DR*09∶0120284A*33∶03-B*58∶0136686B*13∶01-DR*12∶0230201A*33∶03-B*58∶01-DR*03∶0115901A*33∶03-B*44∶0324780B*52∶01-DR*15∶0221468A*02∶01-B*13∶01-DR*12∶0214796A*02∶01-B*13∶0118857B*40∶06-DR*09∶0120889A*33∶03-B*44∶03-DR*13∶0213491A*11∶01-B*40∶0116878B*58∶01-DR*03∶0120855A*33∶03-B*58∶01-DR*13∶0212696A*02∶01-B*15∶1116026B*15∶01-DR*15∶0117280A*02∶07-B*46∶01-DR*08∶039679A*24∶02-B*40∶0115502B*44∶03-DR*07∶0116892A*01∶01-B*57∶01-DR*07∶019612A*02∶01-B*15∶0115405B*44∶03-DR*13∶0215977A*01∶01-B*37∶01-DR*10∶018017A*24∶02-B*40∶0613122B*46∶01-DR*08∶0315954A*33∶03-B*44∶03-DR*07∶017483

3討論

本研究在黑龍江地區(qū)人群中共檢測(cè)到286個(gè)HLA-A、B、DRB1等位基因，進(jìn)一步豐富了中國(guó)人群HLA等位基因的多態(tài)性。其中又以B座位的多態(tài)性最為豐富，不僅是因?yàn)锽座位比A、DRB1座位存在數(shù)目更多的等位基因，還因?yàn)锽座位中常見等位基因并非處于絕對(duì)優(yōu)勢(shì)地位(表2)，其>0.1的常見等位基因累計(jì)頻率為0，明顯

80個(gè)A位點(diǎn)等位基因、136個(gè)B位點(diǎn)等位基因和70個(gè)DR位點(diǎn)等位基因的編碼基因，理論上可組成10880種A-B、9520種B-DRB1、761600種A-B-DRB1單體型，但其中有90%(9793/10880)的A-B、86.04%(8191/9520)的B-DRB1和99.42%(757172/761600)的A-B-DRB1單體型(表3)未觀察到或頻率<10-6，提示黑龍江地區(qū)人群中HLA-A,B,DRB1等位基因間存在連鎖不平衡。這與A-B,B-DRB1和A-B-DRB1單體型的實(shí)際熵值低于理論熵值的計(jì)算結(jié)果(7.55<3.74+5.05,7.84<5.05+4.19,9.86<3.74+5.05+4.19)相符合。黑龍江地區(qū)人群中，分別有267條HLA-A-B單體型和357條HLA-B-DRB1單體型呈現(xiàn)顯著的正連鎖不平衡(ALD>0,HF≥1.09×10-4,χ2>3.84),其中A*02∶53N-B*27∶04、A*25∶01-B*18∶01等12條A-B單體型和B*42∶01-DR*13∶02、B*27∶24-DR*04∶05等18條B-DRB1單體型表現(xiàn)為強(qiáng)連鎖不平衡(RLD>0.80)。在呈現(xiàn)強(qiáng)連鎖不平衡的A-B、B-DRB1單體型中，屬于常見單體型的僅為A*30∶01-B*13∶02和B*13∶02-DR*07:01(表5)。由此可見，高頻單體型不一定連鎖不平衡，頻率較低的等位基因間也存在顯著的連鎖不平衡，如A*02∶53N-B*27∶04、B*42∶01-DR*13∶02等單體型，頻率都<0.001，但在統(tǒng)計(jì)學(xué)角度上表現(xiàn)為強(qiáng)連鎖不平衡，這是本研究調(diào)查樣本量大的結(jié)果。

在黑龍江地區(qū)人群中觀察到158條常見的HLA-A-B-DRB1單體型，其中頻率最高的3條HLA-A-B-DRB1單體型是A*30∶01-B*13∶02-DR*07∶01、A*02∶07-B*46∶01-DR*09∶01、A*33∶03-B*58∶01-DR*03∶01，這與黑龍江地區(qū)漢族低分辨單體型研究結(jié)果一致[4]。黑龍江地區(qū)人群HLA-A-B-DRB1單體型與中國(guó)北方(北京、天津、石家莊、山東半島)漢族人、南方(廣東、上海、臺(tái)灣)漢族人相比，北方、南方漢族人常見的單體型在本組中頻率均>0.001[5-9]，而本組頻率最高的10種HLA-A-B-DRB1單體型中僅A*01∶01-B*57∶01-DR*07∶01單體型在南方漢族人中頻率<0.001 ，其余均>0.001，而且與山東半島漢族人前10種HLA-A-B-DRB1單體型完全相同[6]；與亞洲其他地區(qū)相比，在韓國(guó)、越南人群中常見的HLA-A-B-DRB1單體型在本組中均有檢出[10,11]。在韓國(guó)人群頻率最高的10種HLA-A-B-DRB1單體型中有5種單體型與本組相同，這可能是同屬黃種人且地域較接近的緣故，而其中A*30∶04-B*14∶01-DR*04∶04、A*02∶01-B*27∶05-DR*01∶01在本組中頻率則<0.001，說明兩者有各自的單體型分布特征。與越南人相比，在越南人群中頻率最高的10種HLA-A-B-DRB1單體型中有4條單體型與本組相同，而有4種HLA-A-B-DRB1單體型在本組中頻率則<0.001，說明與越南人的單體型分布特征差異增大；與西方人相比，德國(guó)人群中常見的A29∶02-B44∶03-DR07∶01、A02∶01-B15∶01-DR13∶01、美國(guó)人群中的A29∶02-B44∶03-DR07∶01單體型在本組中均未觀察到[12,13]。通過以上可以看出隨著地域、人種不同，HLA-A-B-DRB1單體型頻率存在明顯差異。

黑龍江省地處中國(guó)的最北端，頻率較高的HLA單體型的分布符合中國(guó)北方漢族人群特征，與中國(guó)漢族人群有較大的相似性。分析HLA-A,B,DRB1等位基因多態(tài)性、單體型頻率及連鎖不平衡的特征可為估計(jì)捐獻(xiàn)者資料的例數(shù)對(duì)患者查詢的滿足程度、與患者查詢的匹配幾率以及與移植成活或預(yù)后的關(guān)系等研究提供依據(jù)。

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[收稿2015-04-21]

(編輯許四平)

[關(guān)鍵詞]HLA-A/B/DRB1；高分辨基因分型；基因/單體型頻率

Polymorphism research of HLA-A,B,DRB1 high resolution alleles and haplotypes in population from Heilongjiang

LIXin,DINGJuan,WANGXin,HOULing,LIUYing，LIUJie.BloodCenterofHaerbin，HeilongjiangProvince,Harbin150056，China

[Abstract]Objective:To research the distribution features of HLA-A,B and DRB1 high resolution alleles and haplotypes in Heilongjiang population.Methods: PCR-SBT methods was applied for HLA-A,B,DRB1 high resolution genotyping of 13 670 unrelated and healthy donors in region of Heilongjiang,and haplotype frequencies were calculated by counting and maximum likelihood method.Results: A total of 286 HLA alleles were observed and the most frequent alleles(>0.1)were A* 02∶01，A*24∶02，A*11∶01，DR*07∶01，DR*09∶01 and DR*15∶01.Among 1 087 kinds of HLA-A-B haplotype ,there were 22 kinds frequency higher than 0.01,and 267 kinds with statistically significant and positive linkage disequilibrium(ALD>0,HF≥1.09×10-4,χ2>3.84).Moreover,among 1 329 kinds of HLA-B-DRB1 haplotype,there were 19 haplotypes frequency higher than 0.01,and 357 kinds with statistically significant and positive linkage disequilibrium.1 348 kinds of A-B-DR Haplotype were informative with frequency ≥1.66×10-4in 4 428 haplotypes,and a total of 17 kinds of A-B-DR haplotype frequency higher than 0.005.Conclusion: Get the distribution features of HLA high resolution allele and haplotype in Heilongjiang population,and associated genetic parameters,Distribution of alleles and haplotypes close to northern Han population,but have their own distribution.

[Key words]HLA-A/B/DRB1；High resolution genotyping；Gene/Haplotype frequency

作者簡(jiǎn)介：李鑫(1978年-)，女，碩士，主管技師，主要從事人類免疫遺傳學(xué)研究，E-mail:xin53@163.com。

通訊作者②，E-mail:liuying54609@126.com；liujie1954@126.com。

中圖分類號(hào)R457.1+1
①本文為黑龍江省衛(wèi)生廳科研課題(No.2012-082)。

文獻(xiàn)標(biāo)志碼A

文章編號(hào)1000-484X(2016)01-0083-07

doi:10.3969/j.issn.1000-484X.2016.01.018