国际输血及血液学杂志
國際輸血及血液學雜誌
국제수혈급혈액학잡지
INTERNATIONAL JOURNAL OF BLOOD TRANSFUSION AND HEMATOLOGY
2012年
2期
101-104
,共4页
人类血小板抗原%流式细胞仪结合序列特异性寡核苷酸探针%基因分型%血小板供者资料库
人類血小闆抗原%流式細胞儀結閤序列特異性寡覈苷痠探針%基因分型%血小闆供者資料庫
인류혈소판항원%류식세포의결합서렬특이성과핵감산탐침%기인분형%혈소판공자자료고
human platelet antigens%FLOW-SSO%genotyping%platelet donor database
目的 探讨高通量流式细胞仪结合序列特异性寡核苷酸探针(FLOW-SSO)技术在人类血小板抗原( HPA)基因分型中的应用.方法 采用FLOW-SSO技术对1378例汉族血小板捐献者HPA1～6,15系统的基因型进行检测；并以10份国际输血协会(ISBT)第12届血小板免疫学术研讨会提供的质控样品对FLOW-SSO技术的可靠性进行考核.以3家供应商提供的不同聚合酶链反应-序列特异引物法(PCR-SSP)试剂,对50份血小板样品的基因分型结果进行比对试验.分型结果采用x2检验考察HPA系统的基因型频率分布是否符合Hardy-Weinberg遗传平衡定律.结果 通过FLOW-SSO方法获得的1379例血小板供者HPA等位基因频率如下,HPA-1a为0.9927,HPA-1b为0.0073,HPA-2a为0.9536,HPA-2b为0.0464,HPA-3a为0.5700,HPA-3b为0.4300,HPA-4a为0.9982,HPA -4b为0.0018,HPA-5a为0.9804,HPA-5b为0.0196,HPA-6a为0.9822,HPA-6b为0.0178,HPA- 15a为0.5337及HPA-15b为0.4663.基因频率检测结果经x2检验,其分布符合Hardy-Weinberg遗传平衡定律.ISBT提供的10份样品考核结果相符率为100％.50份血小板样本采用3种不同的PCR-SSP试剂测定的HPA1～6,15基因分型结果,与FLOW-SSO技术测定结果完全一致.结论 FLOW-SSO基因分型技术有便捷、重复性好及高通量等特点,适用于血小板供者资料库的血小板基因分型.
目的 探討高通量流式細胞儀結閤序列特異性寡覈苷痠探針(FLOW-SSO)技術在人類血小闆抗原( HPA)基因分型中的應用.方法 採用FLOW-SSO技術對1378例漢族血小闆捐獻者HPA1～6,15繫統的基因型進行檢測；併以10份國際輸血協會(ISBT)第12屆血小闆免疫學術研討會提供的質控樣品對FLOW-SSO技術的可靠性進行攷覈.以3傢供應商提供的不同聚閤酶鏈反應-序列特異引物法(PCR-SSP)試劑,對50份血小闆樣品的基因分型結果進行比對試驗.分型結果採用x2檢驗攷察HPA繫統的基因型頻率分佈是否符閤Hardy-Weinberg遺傳平衡定律.結果 通過FLOW-SSO方法穫得的1379例血小闆供者HPA等位基因頻率如下,HPA-1a為0.9927,HPA-1b為0.0073,HPA-2a為0.9536,HPA-2b為0.0464,HPA-3a為0.5700,HPA-3b為0.4300,HPA-4a為0.9982,HPA -4b為0.0018,HPA-5a為0.9804,HPA-5b為0.0196,HPA-6a為0.9822,HPA-6b為0.0178,HPA- 15a為0.5337及HPA-15b為0.4663.基因頻率檢測結果經x2檢驗,其分佈符閤Hardy-Weinberg遺傳平衡定律.ISBT提供的10份樣品攷覈結果相符率為100％.50份血小闆樣本採用3種不同的PCR-SSP試劑測定的HPA1～6,15基因分型結果,與FLOW-SSO技術測定結果完全一緻.結論 FLOW-SSO基因分型技術有便捷、重複性好及高通量等特點,適用于血小闆供者資料庫的血小闆基因分型.
목적 탐토고통량류식세포의결합서렬특이성과핵감산탐침(FLOW-SSO)기술재인류혈소판항원( HPA)기인분형중적응용.방법 채용FLOW-SSO기술대1378례한족혈소판연헌자HPA1～6,15계통적기인형진행검측；병이10빈국제수혈협회(ISBT)제12계혈소판면역학술연토회제공적질공양품대FLOW-SSO기술적가고성진행고핵.이3가공응상제공적불동취합매련반응-서렬특이인물법(PCR-SSP)시제,대50빈혈소판양품적기인분형결과진행비대시험.분형결과채용x2검험고찰HPA계통적기인형빈솔분포시부부합Hardy-Weinberg유전평형정률.결과 통과FLOW-SSO방법획득적1379례혈소판공자HPA등위기인빈솔여하,HPA-1a위0.9927,HPA-1b위0.0073,HPA-2a위0.9536,HPA-2b위0.0464,HPA-3a위0.5700,HPA-3b위0.4300,HPA-4a위0.9982,HPA -4b위0.0018,HPA-5a위0.9804,HPA-5b위0.0196,HPA-6a위0.9822,HPA-6b위0.0178,HPA- 15a위0.5337급HPA-15b위0.4663.기인빈솔검측결과경x2검험,기분포부합Hardy-Weinberg유전평형정률.ISBT제공적10빈양품고핵결과상부솔위100％.50빈혈소판양본채용3충불동적PCR-SSP시제측정적HPA1～6,15기인분형결과,여FLOW-SSO기술측정결과완전일치.결론 FLOW-SSO기인분형기술유편첩、중복성호급고통량등특점,괄용우혈소판공자자료고적혈소판기인분형.
Objective To explore the application of the high-throughput flow-sequence specific oligonucleotide probes (FLOW-SSO) technology in human platelet antigen (HPA) genotyping. Method A total of 1378 platelet donors of Han ethnic group were genotyped by FLOW-SSO for HPA system 1-6 and 15 Ten reference samples provided from 12th platelet immunology workshop of International Society of Blood Transfusion (ISBT) were tested as control group, and 50 samples were detected by the PCR-SSP with 3 different manufacturing sources reagents as the comparison test. x2 test was used to study the genotype frequencies of the HPA system, whether conformed the distribution in Hardy-Weinberg genetic equilibrium. Results The distribution of the HPA allelic frequencies of the 1378 donors were as follows:HPA-1a 0.9927 and HPA-1b 0.0073, HPA-2a 0.9536 and HPA-2b 0.0464, HPA-3a 0.5700 and HPA-3b 0.4300, HPA-4a 0.9982 and HPA-4b 0.0018, HPA-5a 0.9804 and HPA-5b 0.0196, HPA-6a 0.9822 and HPA-6b0.0178, HPA-15a 0.5337 and HPA-15b0.4663, which were in agreement with the Hardy-Weinberg genetic equilibrium. Ten reference samples of quality controls were 100％ in concordance with results of ISBT. The 50 genotypes were all matched in comparison of the methods between FLOW-SSO and PCR-SSP. Conclusions With the advantages of rapid, repeatable and high-throughput testing, the streamlined FLOWSSO technology for HPA system 1-6 and 15 genotyping was applicable to large-scare genetic population studies.
