Analysis for Eight ABO Alleles in Korean Population

Sang Hoon Song; Ho Eun Chang; Kwang Chul Ryu; Hyun Jung Lee; Kyoung Un Park; Junghan Song; Kyou Sup Han

doi:10.3343/kjlm.2006.26.5.374

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Song, Chang, Ryu, Lee, Park, Song, and Han: Analysis for Eight ABO Alleles in Korean Population

Original Article

Korean J Leg Med 2006;26(5):374-379.

Published online: 10 February 2006

DOI: https://doi.org/10.3343/kjlm.2006.26.5.374

Analysis for Eight ABO Alleles in Korean Population

Sang Hoon Song, M.D.¹, Ho Eun Chang, M.T.², Kwang Chul Ryu, M.T.², Hyun Jung Lee, M.T.², Kyoung Un Park, M.D.^1,², Junghan Song, M.D.^1,², Kyou Sup Han, M.D.¹

¹Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea

²Seoul National University Bundang Hospital, Bundang, Korea

Received 1 June 2006 Revised 25 August 2006 Accepted 25 August 2006

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

ABO genotyping is a useful tool in case of ABO discrepancies and in legal medicine. Recent knowledge of various alleles in the ABO gene has led to the need of a different method that can cover numerous polymorphisms. We performed a polymerase chain reaction using sequence-specific priming (PCR-SSP) with 12 primer sets and evaluated its value in the detection of 8 ABO alleles.

Methods

Genomic DNA was isolated from peripheral blood of 222 unrelated Koreans. Sequence-specific primer sets for the nucleotides 261, 297, 467, 802, 803, and 1059 were selected, and 12 PCR reactions were performed for each sample. Direct sequencing was performed to evaluate the accuracy of discrimination between A¹(Pro) and A¹(Leu) and between O¹ and O^1v.

Results

All the ABO genotype patterns were in an exact match with the ABO phenotypes. The results from sequencing and PCR-SSP were equivalent. The allele frequencies of A¹, B, O¹, and O^1v were 27.25%, 19.82%, 27.25%, and 25.68% respectively. Out of total 121 A¹ alleles, 6 (4.96%) were A¹(Pro) alleles and 115 (95.04%) were A¹(Leu) alleles. No A², O², or CisAB alleles were found in this study.

Conclusions

The proportion of O^1v allele was similar to that of O¹ allele. This was an unexpected result. We developed a method for detecting 8 ABO alleles by PCR-SSP; the method was accurate and was able to discriminate between A¹(Pro) and A¹(Leu) and between O¹ and O^1v.

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Keywords: ABO, Polymerase chain reaction, Allele, A¹(Leu), O^1v

References

1. Yamamoto F, Clausen H, White T, Marken J, Hakomori S. Molecular genetic basis of the histo-blood group ABO system. Nature. 1990; 345:229–33.

2. Gassner C, Schmarda A, Nussbaumer W, Schonitzer D. ABO glycosyltransferase genotyping by polymerase chain reaction using sequence-specific primers. Blood. 1996; 88:1852–6.

3. Seltsam A, Hallensleben M, Kollmann A, Burkhart J, Blasczyk R. Systematic analysis of the ABO gene diversity within exons 6 and 7 by PCR screening reveals new ABO alleles. Transfusion. 2003; 43:428–39.

4. Lee JC, Chang JG. ABO genotyping by polymerase chain reaction. J Forensic Sci. 1992; 37:1269–75.

5. Fukumori Y, Ohnoki S, Shibata H, Yamaguchi H, Nishimukai H. Genotyping of ABO blood groups by PCR and RFLP analysis of 5 nucleotide positions. Int J Legal Med. 1995; 107:179–82.

6. Oh HB, Cho YJ, Cho YH, Hwang YS, Kim DS, Kim SI. Genotyping of ABO and D antigens. Korean J Blood Transfus. 1997; 8:31–7.

7. Kang SH, Shin DH, Cho HC, Lee KM, Han KS. Simple ABO genotyping method using three polymorphic sites at the ABO locus. Korean J Blood Transfus. 1998; 9:155–65.

8. Kim MJ, Shin JW, Kim YH, Kim HO, Cho SR, Kim WJ. Application of ABO genotyping in determination of ABO subgroups. Korean J Blood Transfus. 1998; 9:209–17.

9. Yip SP. Single-tube multiplex PCR-SSCP analysis distinguishes 7 common ABO alleles and readily identifies new alleles. Blood. 2000; 95:1487–92.

10. Olsson ML, Irshaid NM, Hosseini-Maaf B, Hellberg A, Moulds MK, Sareneva H, et al. Genomic analysis of clinical samples with serologic ABO blood grouping discrepancies: identification of 15 novel A and B subgroup alleles. Blood. 2001; 98:1585–93.

11. Ferri G, Bini C, Ceccardi S, Pelotti S. ABO genotyping by minisequencing analysis. Transfusion. 2004; 44:943–4.

12. Doi Y, Yamamoto Y, Inagaki S, Shigeta Y, Miyaishi S, Ishizu H. A new method for ABO genotyping using a multiplex single-base primer extension reaction and its application to forensic casework samples. Leg Med (Tokyo). 2004; 6:213–23.

13. Ferri G, Bini C, Ceccardi S, Ingravallo F, Lugaresi F, Pelotti S. Minisequencing-based genotyping of duffy and ABO blood groups for forensic purposes. J Forensic Sci. 2006; 51:357–60.

14. Seo DH, Kim SY, Kin JY, Park SS, Lee JB, Han KS. Molecular characteristics of B subgroups in Koreans. Korean J Lab Med. 2005; 25:280–4.

15. Ringel PF, Weiler G, Bein G. Errors in ABO typing of blood stains using PCR. Int J Legal Med. 2000; 113:352–5.

16. Kang SH, Fukumori Y, Ohnoki S, Shibata H, Han KS, Nishimukai H, et al. Distribution of ABO genotypes and allele frequencies in a Korean population. Jpn J Hum Genet. 1997; 42:331–5.

17. Olsson ML, Santos SE, Guerreiro JF, Zago MA, Chester MA. Heterogeneity of the O alleles at the blood group ABO locus in Amerindians. Vox Sang. 1998; 74:46–50.

18. Chester MA, Olsson ML. The ABO blood group gene: a locus of considerable genetic diversity. Transfus Med Rev. 2001; 15:177–200.

19. Yip SP. Sequence variation at the human ABO locus. Ann Hum Genet. 2002; 66:1–27.

20. Fukumori Y, Ohnoki S, Shibata H, Nishimukai H. Suballeles of the ABO blood group system in a Japanese population. Hum Hered. 1996; 46:85–91.

21. Nishimukai H, Fukumori Y, Okiura T, Yuasa I, Shinomiya T, Ohnoki S, et al. Genotyping of the ABO blood group system: analysis of nucleotide position 802 by PCR-RFLP and the distribution of ABO genotypes in a German population. Int J Legal Med. 1996; 109:90–3.

22. Cho D, Shin MG, Yazer MH, Kee SJ, Shin JH, Suh SP, et al. The genetic and phenotypic basis of blood group A subtypes in Koreans. Transfus Med. 2005; 15:329–34.

23. Irshaid NM, Chester MA, Olsson ML. Allele-related variation in minisatellite repeats involved in the transcription of the blood group ABO gene. Transfus Med. 1999; 9:219–26.

24. Suzuki K, Iwata M, Tsuji H, Takagi T, Tamura A, Ishimoto G, et al. A de novo recombination in the ABO blood group gene and evidence for the occurrence of recombination products. Hum Genet. 1997; 99:454–61.

25. Olsson ML, Chester MA. A rapid and simple ABO genotype screening method using a novel B/O2 versus A/O2 discriminating nucleotide substitution at the ABO locus. Vox Sang. 1995; 69:242–7.

26. Suzuki K. ABO blood group alleles and genetic recombination. Leg Med (Tokyo). 2005; 7:205–12.

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Table 1.

Oligonucleotide primers used in typing of 8 ABO alleles

Polymorphic site	Name	Primer sequence (5′to 3′)
Nucleotide 261	O1-F	GGA AGG ATG TCC TCG TGG TA
	nO1-F	GGA AGG ATG TCC TCG AGG TG
	O1-R	ATA TAT ATG GCA AAC ACA GTT AAC CCA ATG
Nucleotide 802	O2B-F	AGT GGA CGT GGA CAT GGA GTT CC
	O2-R	TCG ACC CCC CGA AGA AGC T
	nO2-R	CGA CCC CCC GAA GAA GCC
Nucleotide 803	O2B-F	AGT GGA CGT GGA CAT GGA GTT CC
	B-R	ATC GAC CCC CCG AAG AGC G
	nB-R	CCG ACC CCC CGA AGA GCC
Nucleotide 1059	A2-F	GAG GCG GTC CGG AAG CG
	nA2-F	GAG GCG GTC CGG AAC ACG
	A2-R	GGG TGT GAT TTG AGG TGG GGA C
Nucleotide 297	297G-F	CAT TGT CTG GGA GGG CAC G
	297A-F	CAT TGT CTG GGA GGG CAC A
	297-R	CTT GAT GGC AAA CAC AGT TAA C
Nucleotide 467	467-F	ACG TGG CTT TCC TGA AGC TG
	467T-R	GCG GAG CAC CGC GGC CG
	467C-R	GCG GGG CAC CGC GGC CA
Internal control	HGH-F	TGC CTT TTT AAC CAT TCC CTT A
	HGH-R	CCA CTC ACG GAT TTC TGT TGT GTT TC

Artificially changed base pairs are expressed as underlined.

Table 2.

Possible 8 alleles matched by polymorphisms on six nucleotide positions at the ABO locus

Nucleotide position						Allele
261	802	803	1059	297	467	Allele
G	G	G	C	A	C	A¹(Pro)
G	G	G	C	A	T	A¹(Leu)
G	G	G	Δ	A	T	A²
G	G	C	C	G	C	B
Δ	G	G	C	A	C	O¹
G	A	G	C	G	C	O²
G	G	C	C	A	T	CisAB
Δ	G	G	C	G	C	O1V

Abbreviation: Δ, deletion.

Table 3.

Possible patterns detected with PCR-SSP and results of ABO genotyping in 222 Korean blood donors

261	261	802	803	803	803	1059	1059	297	297	467	467	Genotype	Pheno-type	Number (%)
1	2	3	4	5	6	7	8	9	10	11	12
01	NonO1	O2	NonO2	B	NonB	A2	NonA2
Δ	G	A	G	C	G	Δ	C	G	A	T	C
+			+		+		+		+		+	O¹/O¹	O	20 (9.01)
+	+	+	+		+		+	+	+		+	O¹/O²	O	0 (0.00)
+	+		+	+	+		+	+	+		+	O¹/B	B	20 (9.01)
+	+		+		+		+		+	+/-	+	O¹/A¹	A₁	33 (14.86)
+	+		+		+	+	+		+	+	+	O¹/A²	A₂	0 (0.00)
	+	+			+		+	+			+	O²/O²	O	0 (0.00)
	+	+	+	+	+		+	+			+	O²/B	B	0 (0.00)
	+	+	+		+		+	+	+	+/-	+	O²/A¹	A₁	0 (0.00)
	+	+	+		+	+	+	+	+	+	+	O²/A²	A₂	0 (0.00)
	+		+	+			+	+			+	B/B	B	8 (3.60)
	+		+	+	+		+	+	+	+/-	+	B/A¹	A₁B	27 (12.16)
	+		+	+	+	+	+	+	+	+	+	B/A²	A₂B	0 (0.00)
	+		+		+	+	+		+	+/-	+	A²/A¹	A₁	0 (0.00)
	+		+		+	+			+	+		A²/A²	A₂	0 (0.00)
	+		+		+		+		+	+/-	+/-	A¹/A¹	A₁	12 (5.41)
+			+		+		+	+	+		+	O^1v/O¹	O	28 (12.61)
+	+		+		+		+	+	+	+/-	+	O^1v/A¹	A₁	37 (16.67)
+	+		+	+	+		+	+			+	O^1v/B	B	25 (11.26)
+			+		+		+	+			+	O^1v/O^1v	O	12 (5.41)

Abbreviations: PCR-SSP, polymerase chain reaction using sequence-specific priming; Δ, deletion.

Table 4.

ABO genotype frequencies (%) in this study and comparison with other ethnic groups

Genotype	This study (N=222)	Korean [16] (N=253)	Japanese [20] (N=520)	Chinese [9] (N=125)	German [21] (N=169)	European [9] (N=98)
A¹(Leu)/A¹(Leu)	4.95	4.3	4.2	2.4	0.0^†	1.02
A¹(Pro)/A¹(Pro)	0.00	0.4	0.8	0.0	4.7^†	1.02
A¹(Leu)/A¹(Pro)	0.45	0.4	2.5	0.8	3.5^*	0.00
A¹(Leu)/O¹	14.41	14.2	11.9	18.4	6.5^†	0.00^†
A¹(Pro)/O¹	0.45	2.4	4.6	0.0	17.2^†	14.29^†
A¹(Leu)/O¹^v	15.32	8.3^*	12.1	5.6^†	3.5^†	0.00^†
A¹(Pro)/O¹^v	1.35	0.4	3.3	0.0	11.2^†	3.06
B/B	3.60	4.3	2.5	0.8	0.6^*	2.04
B/O¹	9.01	15.0^*	10.4	14.4	2.4^†	9.18
B/O¹^v	11.26	7.5	9.6	9.6	2.4^†	4.08^*
O¹/O¹	9.01	13.4	6.3	12.0	19.0^†	15.31
O¹/O¹^v	12.61	13.4	15.0	20.0	19.0	17.35
O¹^v/O¹^v	5.41	5.2	6.2	6.4	3.5	2.04
A¹(Leu)/B	11.71	10.3	8.3	6.4	1.2^†	1.02^†
A¹(Pro)/B	0.45	0.4	2.3	1.6	1.2	4.08^*
Others^‡	−	−	−	1.6	4.1	25.51

^* P<0.05 and

^† P<0.01 as compared to this study.

^‡ Others include genotypes with A², O², or other O alleles other than O¹ or O^1v.

Table 5.

ABO allele frequencies in various populations

Population	A¹(Leu)	A¹(Pro)	B	O¹	O1v	O²	A²	Others^‡
This study	0.259	0.013	0.198	0.273	0.257	0.000	0.000	−
Korean[16]	0.209	0.022	0.209	0.360^†	0.200^*	0.000	Not tested	−
Japanese[20]	0.216	0.071^†	0.178	0.273	0.262	0.000	Not tested	−
Chinese[9]	0.180^*	0.012	0.168	0.384^†	0.248	0.000	0.000	0.008
German[21]	0.077^†	0.213^†	0.047^†	0.426^†	0.216	0.021	Not tested	−
European[9]	0.015^†	0.138^†	0.128^*	0.408^†	0.163^†	0.010^*	0.056^†	0.082

^* P<0.05 and

^† P<0.01 as compared to this study.

^‡ Others are rare A or Oalleles with unexpected base change or deletion not defined in our method.

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