• 四川大學(xué)華西醫(yī)院消化外科實(shí)驗(yàn)室(成都610041);

【摘要】目的采用單管PCR擴(kuò)增(ST Amp)方法建立高效的人類白細(xì)胞抗原-DRB1（HLA-DRB1）基因測(cè)序分型(SBT)技術(shù)并探討其應(yīng)用價(jià)值。
方法合成7個(gè)5′端特異性擴(kuò)增引物和1個(gè)3′端共用引物，將上述引物混合在一起組成單個(gè)PCR反應(yīng)用于HLA-DRB1的DNA測(cè)序分型分析。
結(jié)果所有樣本都能成功分型，分型結(jié)果準(zhǔn)確可靠，重復(fù)性好。
結(jié)論ST Amp法改進(jìn)了測(cè)序分型技術(shù)，具有高分辨率、高特異性和高效率的特點(diǎn)，值得推廣應(yīng)用。

引用本文： 陳耿臻,周總光,鄭雪蓮,周斌,辜俊. PCR-ST Amp法應(yīng)用于HLA-DRB1測(cè)序分型的研究. 中國(guó)普外基礎(chǔ)與臨床雜志, 2005, 12(3): 258-260. doi: 復(fù)制

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6. GarciaSanchez F, FernandezDeVelasco J, Lillo R, et al. Two new HLA class I alleles recognised by PCR sequencespecific primer and sequencing based typing: B*3805 and Cw*0408 [Ｊ]. Tissue Antigens, 2002; 59(1)∶47.
7. EllexsonTurner ME, Leffell MS, Zachary AA, et al. Sequencebased typing of HLA class I alleles in Alaskan Yupik Eskimo [Ｊ]. Hum Immunol, 2001; 62(6)∶639.
8. AlHussein KA, Rama NR, Butt AI, et al. HLA class II sequencebased typing in normal Saudi individuals [Ｊ]. Tissue Antigens, 2002; 60(3)∶259.
9. Curcio M, Lapi S, Italia S, et al. Identification of a novel HLADPB1 allele, DPB1*02014, by sequencebased typing [Ｊ]. Tissue Antigens, 2002; 59(1)∶58.
10. Adams S, Krausa P, McGinnis M, et al. Practicality of high throughput HLA sequencingbased typing [C]. ASHI Quarterley, 2001∶54-57.
11. Sayer D, Whidborne R, Brestovac B, et al. HLADRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors [Ｊ]. Tissue Antigens， 2001; 57(1)∶46.
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  2. 2. 李成濤. HLA基因分型技術(shù)的進(jìn)展與展望 [Ｊ]. 法醫(yī)學(xué)雜志， 2004； 20(2)∶120.
  3. 3. Sayer DC, Whidborne R, De Santis D, et al. A multicenter international evaluation of singletube amplification protocols for sequencingbased typing of HLADRB1 and HLADRB3,4,5 [Ｊ]. Tissue Antigens, 2004; 63(5)∶412.
  4. 4. Madrigal JA, Arguello R, Scott I, et al. Molecular histocompatibility typing in unrelated donor bone marrow transplantation [Ｊ]. Blood Rev, 1997; 11(2)∶105.
  5. 5. Hao GQ, Xiao LL. The application of molecular biology techniques in HLAtyping [Ｊ]. Zhongguo Shi Yan Xue Ye Xue Za Zhi， 2002; 10(3)∶268.
  6. 6. GarciaSanchez F, FernandezDeVelasco J, Lillo R, et al. Two new HLA class I alleles recognised by PCR sequencespecific primer and sequencing based typing: B*3805 and Cw*0408 [Ｊ]. Tissue Antigens, 2002; 59(1)∶47.
  7. 7. EllexsonTurner ME, Leffell MS, Zachary AA, et al. Sequencebased typing of HLA class I alleles in Alaskan Yupik Eskimo [Ｊ]. Hum Immunol, 2001; 62(6)∶639.
  8. 8. AlHussein KA, Rama NR, Butt AI, et al. HLA class II sequencebased typing in normal Saudi individuals [Ｊ]. Tissue Antigens, 2002; 60(3)∶259.
  9. 9. Curcio M, Lapi S, Italia S, et al. Identification of a novel HLADPB1 allele, DPB1*02014, by sequencebased typing [Ｊ]. Tissue Antigens, 2002; 59(1)∶58.
  10. 10. Adams S, Krausa P, McGinnis M, et al. Practicality of high throughput HLA sequencingbased typing [C]. ASHI Quarterley, 2001∶54-57.
  11. 11. Sayer D, Whidborne R, Brestovac B, et al. HLADRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors [Ｊ]. Tissue Antigens， 2001; 57(1)∶46.