【Abstract】ObjectiveTo develop a new singletube polymerase chain reaction amplification (ST Amp) protocol for the efficient sequencing-based typing (SBT) of human leukocyte antigen DRB1（HLA-DRB1）.
MethodsA set of 7 group-specific exonic 5′ amplification primers and a single generic 3′ primer were included together in a single PCR mix to facilitate a single PCR amplification per sample for HLA-DRB1 typing.
ResultsAll samples were successfully typed, the typing result was accurate and repeatable.
ConclusionST Amp technique has resulted in the ability to perform high-resolution, high-specificity and high-throughput HLA-DRB1 typing by DNA sequencing.
Citation：
CHEN Gengzhen,ZHOU Zongguang,ZHENG Xuelian,ZHOU Bin,GU Jun.. Single-Tube Polymerase Chain Reaction Amplification Protocol for SequencingBased Typing of Human Leukocyte Antigen DRB1. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2005, 12(3): 258-260. doi:
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Curcio M, Lapi S, Italia S, et al. Identification of a novel HLADPB1 allele, DPB1*02014, by sequencebased typing [Ｊ]. Tissue Antigens, 2002; 59(1)∶58.
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Sayer D, Whidborne R, Brestovac B, et al. HLADRB1 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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- 1. 谭建明，周永昌，唐孝达. 组织配型技术与临床应用 [M]. 第1版. 北京： 人民卫生出版社， 2002∶ 12~22.
- 2. 李成涛. HLA基因分型技术的进展与展望 [Ｊ]. 法医学杂志， 2004； 20(2)∶120.
- 3. Sayer DC, Whidborne R, De Santis D, et al. A multicenter international evaluation of singletube amplification protocols for sequencingbased typing of HLADRB1 and HLADRB3,4,5 [Ｊ]. Tissue Antigens, 2004; 63(5)∶412.
- 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. Hao GQ, Xiao LL. The application of molecular biology techniques in HLAtyping [Ｊ]. Zhongguo Shi Yan Xue Ye Xue Za Zhi， 2002; 10(3)∶268.
- 6. GarciaSanchez F, FernandezDeVelasco J, Lillo R, et al. Two new HLA class I alleles recognised by PCR sequencespecific primer and sequencing based typing: B*3805 and Cw*0408 [Ｊ]. Tissue Antigens, 2002; 59(1)∶47.
- 7. EllexsonTurner ME, Leffell MS, Zachary AA, et al. Sequencebased typing of HLA class I alleles in Alaskan Yupik Eskimo [Ｊ]. Hum Immunol, 2001; 62(6)∶639.
- 8. AlHussein KA, Rama NR, Butt AI, et al. HLA class II sequencebased 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 HLADPB1 allele, DPB1*02014, by sequencebased typing [Ｊ]. Tissue Antigens, 2002; 59(1)∶58.
- 10. Adams S, Krausa P, McGinnis M, et al. Practicality of high throughput HLA sequencingbased typing [C]. ASHI Quarterley, 2001∶54-57.
- 11. Sayer D, Whidborne R, Brestovac B, et al. HLADRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors [Ｊ]. Tissue Antigens， 2001; 57(1)∶46.
