ABSTRACT
OBJECTIVES: Human leukocyte antigen (HLA) B27 is a susceptibility allele of ankylosing spondylitis (AS), and HLA-B27 antigen typing is an important indicator for clinical diagnosis of AS, but current typing methods such as sequence specific primer polymerase chain reaction (PCR-SSP) still possess limitation. Therefore, this study aims to analyze the correlation between B27 subtypes and susceptibility to AS in Hunan Province by applying high-resolution polymerase chain reaction-sequence-based typing (PCR-SBT). METHODS: Peripheral blood of 116 patients with suspected AS (suspected AS group) and 121 healthy volunteers (control group) admitted to the Second Xiangya Hospital from January 2020 to December 2020 were collected for HLA-B genotyping by PCR-SBT. Among the patients in the suspected AS group, 23 patients were finally diagnosed with AS (confirmed AS group), and the remaining 93 undiagnosed patients served as the non-confirmed AS group. PCR-SBT and PCR-SSP were used to detect HLA-B27 typing in 116 patients with suspected AS, and the results of the 2 methods were compared. RESULTS: The HLA-B27 allele frequency in the suspected AS group was significantly higher than that in the control group [11.63% vs 2.48%; P<0.001, odds ratio (OR)=5.18, 95% confidence interval (CI) 2.097 to 12.795]. B*27:04, B*27:05, B*27:06, and B*27:07 were detected in the suspected AS group and the control group. The frequency of the B*27:04 allele in the suspected AS group was significantly higher than that in the control group (9.48% vs 1.24%; P<0.001, OR=8.346, 95% CI 2.463 to 28.282). The positive rate of B27 in the suspected AS group and the confirmed AS group (B27+/+ and B27+/-) was significantly higher than that in the control group (χ2=16.579, P<0.001; χ2=94.582, P<0.001, respectively). Among the confirmed AS group, 21 were HLA-B27 carriers, and the B27 positive rate in the confirmed AS group was 91.3%. PCR-SBT could achieve high resolution typing of the HLA-B gene locus, with higher sensitivity, specificity, positive predictive value, negative predictive value, and accuracy than PCR-SSP. CONCLUSIONS: PCR-SBT typing analysis shows a strong correlation between HLA-B * 27:04 and AS in Hunan province. The PCR-SBT method can be used as the preferred option for the auxiliary diagnosis of clinical AS.
Subject(s)
HLA-B27 Antigen , Spondylitis, Ankylosing , Humans , HLA-B27 Antigen/genetics , Spondylitis, Ankylosing/genetics , Genetic Predisposition to Disease , Genetic Testing , Gene FrequencyABSTRACT
HLA-B*46:83 differs from HLA-B*46:03 by one single nucleotide substitution at position 539G>T.
Subject(s)
HLA-B Antigens , Alleles , Base Sequence , China , Exons/genetics , HLA-B Antigens/genetics , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
HLA-DRB1*04:05:23 differs from HLA-DRB1*04:05:01:01 by a single nucleotide substitution at position 234 G > A.
Subject(s)
Alleles , Base Sequence , Exons/genetics , HLA-DRB1 Chains/genetics , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
Compared with HLA-DQB1*03: 01:01:01, the alleles HLA-DQB1*03:432, HLA-DQB1*03:454 and HLA-DQB1*03:465 each show one nucleotide substitution.
Subject(s)
High-Throughput Nucleotide Sequencing , Alleles , Base Sequence , China , HLA-DQ beta-Chains/genetics , HumansABSTRACT
HLA-A*01:348 differs from HLA-A*01:01:01:01 by a single nucleotide substitution at position 140 T > C.
Subject(s)
HLA-A Antigens , Alleles , Base Sequence , China , HLA-A Antigens/genetics , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
HLA-DPB1*1273:01 differs from HLA-DPB1*05:01:01:01 by two nucleotide substitutions in exon 1.
Subject(s)
Alleles , Base Sequence , HLA-DP beta-Chains/genetics , Humans , Polymerase Chain ReactionABSTRACT
Compared with HLA-C*07:02:01:01, the alleles HLA-C*07:900 and HLA-C*07:906 each show one nucleotide substitution respectively.
Subject(s)
HLA-C Antigens , Alleles , HLA-C Antigens/genetics , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
BACKGROUND: Nucleotide mutations in the ABO gene may reduce the activity of glycosyltransferase, resulting in lower levels of A or B antigen expression in red blood cells. Six known splice sites have been identified according to the database of red cell immunogenetics and the blood group terminology of the International Society of Blood Transfusion. Here, we describe six distinct splice site variants in individuals with ABO subtypes. METHODS: The ABO phenotype was examined using a conventional serological method. A polymerase chain reaction sequence-based typing method was used to examine the whole coding sequence of the ABO gene. The ABO gene haplotypes were studied using allele-specific primer amplification or cloning technology. In silico analytic tools were used to assess the functional effect of splice site variations. RESULTS: Six distinct variants in the ABO gene splice sites were identified in nine individuals with ABO subtypes, including c.28 + 1_2delGT, c.28 + 5G > A, c.28 + 5G > C, c.155 + 5G > A, c.204-1G > A and c.374 + 5G > A. c.28 + 1_2delGT was detected in an Aw individual, while c.28 + 5G > A, c.28 + 5G > C, and c.204-1G > A were detected in Bel individuals. c.155 + 5G > A was detected in one B3 and two AB3 individuals, whereas c.374 + 5G > A was identified in two Ael individuals. Three novel splice site variants (c.28 + 1_2delGT, c.28 + 5G > A and c.28 + 5G > C) in the ABO gene were discovered, all of which resulted in low antigen expression. In silico analysis revealed that all variants had the potential to alter splice transcripts. CONCLUSIONS: Three novel splice site variations in the ABO gene were identified in Chinese individuals, resulting in decreased A or B antigen expression and the formation of ABO subtypes.
Subject(s)
ABO Blood-Group System , ABO Blood-Group System/genetics , Alleles , Humans , Mutation/genetics , Phenotype , Sequence Analysis, DNAABSTRACT
Compared with HLA-A*24:02:01:01, the alleles HLA-A*24:02:129 and HLA-A*24:02:135 each show one nucleotide substitution, respectively.
Subject(s)
Polymorphism, Single Nucleotide , Alleles , HLA-A24 Antigen , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
KIR3DL2*00711 differs from KIR3DL2*0070101 by a single nucleotide substitution at position 1344G>A.
Subject(s)
Asian People , Receptors, KIR3DL2 , Alleles , Asian People/genetics , China , Humans , Polymerase Chain Reaction , Receptors, KIR3DL2/genetics , Sequence Analysis, DNAABSTRACT
HLA-DQB1*03:282N differs from HLA-DQB1*03:03:02:01 by a single nucleotide deletion at position 258.
Subject(s)
Alleles , Base Sequence , China , HLA-DQ beta-Chains/genetics , Histocompatibility Testing , Humans , Sequence Analysis, DNAABSTRACT
Compared with HLA-A*11:01:01:01, the alleles HLA-A*11:01:89, -A*11:01:96 and -A*11:01:01:14 each show one nucleotide substitution.
Subject(s)
Asian People , Nucleotides , Alleles , Asian People/genetics , Base Sequence , China , HLA-A Antigens/genetics , Histocompatibility Testing , Humans , Polymorphism, Single Nucleotide , Sequence Analysis, DNAABSTRACT
Compared with HLA-C*01:02:01:01, the alleles HLA-C*01:02:56 and HLA-C*01:02:57 each show one synonymous nucleotide substitution.
Subject(s)
HLA-C Antigens , Alleles , Base Sequence , Exons/genetics , HLA-C Antigens/genetics , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
HLA-DRB1*04:305 differs from HLA-DRB1*04:05:01:01 by two nucleotide substitutions.
Subject(s)
Leukemia , Alleles , Base Sequence , China , HLA-DRB1 Chains/genetics , Humans , Leukemia/genetics , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
The new allele HLA-B*13:109 differs from HLA-B*13:01:01:01 by one nucleotide substitution in exon 4.
Subject(s)
Genes, MHC Class I , HLA-B Antigens , Alleles , Base Sequence , HLA-B Antigens/genetics , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
HLA-DRB1*09:40 differs from HLA-DRB1*09:01:02:01 by one nucleotide substitution at position 214(G>A).
Subject(s)
HLA-DRB1 Chains , Alleles , Base Sequence , China , HLA-DRB1 Chains/genetics , Histocompatibility Testing , Humans , Polymerase Chain ReactionABSTRACT
HLA-C*15:160N differs from HLA-C*15:02:01:01 by a single nucleotide substitution at codon 244 in exon 4.
Subject(s)
Genes, MHC Class I , HLA-C Antigens , Alleles , HLA-C Antigens/genetics , Histocompatibility Testing , Humans , Polymorphism, Single Nucleotide , Sequence Analysis, DNAABSTRACT
HLA-A*02:787 differs from HLA-A*02:07:01 by a single nucleotide substitution at position 278 C > T.
Subject(s)
Asian People , HLA-A Antigens , Alleles , Base Sequence , HLA-A Antigens/genetics , Histocompatibility Testing , Humans , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
The new allele KIR3DL2*113 showed one nucleotide difference with KIR3DL2*0020101 at codon 345.
Subject(s)
Receptors, KIR3DL2 , Alleles , Humans , Receptors, KIR3DL2/genetics , Sequence Analysis, DNAABSTRACT
KIR3DL2*114 differs from KIR3DL2*016 by a single nucleotide substitution at position 329 G>A.
