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Background & objectives: In multitransfused thalassaemic patients, haemagglutination fails to phenotype the patient's blood group antigens due to the presence of donor-derived erythrocytes. DNA-based methods can overcome the limitations of haemagglutination and can be used to determine the correct antigen profile of these patients. This will facilitate the procurement of antigen-matched blood for transfusion to multitransfused patients. Thus, the aim of this study was to compare the serological phenotyping of common and clinically important antigens of Rh, Duffy, Kell, Kidd and MNS blood group systems with molecular genotyping amongst multitransfused thalassaemic patients. Methods: Blood samples from 200 patients with thalassaemia and 100 'O' group regular blood donors were tested using standard serological techniques and polymerase chain reaction-based methods for common antigens/alleles (C, c, D, E, e, Fya, Fyb, Jka, Jkb, K, k, M, N, S, s). Results: Genotyping and phenotyping results were discordant in 77 per cent of thalassaemic patients for five pairs of antithetical antigens of Rh, Duffy, Kell and Kidd blood group systems. In the MNS blood group system, 59.1 per cent of patients showed discrepancy. The rate of alloimmunization among thalassaemics was 7.5 per cent. Interpretation & conclusions: Molecular genotyping enabled the determination of the actual antigen profile in multitransfused thalassaemia patients. This would help reduce the problem of alloimmunization in such patients and would also aid in the better management of transfusion therapy.
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Objective To investigate the killer cell immunoglobulin-like receptor (KIR) genes, KIR2DS4 and its variant KIR1D for an association with syphilis in the comparison between syphilis patients and unrelated healthy subjects. Methods One hundred and ninety syphilis patients and 192 unrelated healthy subjects were performed to determine the KIR genotypes by PCR-SSP method. The gene frequencies of KIR2DS4 and KIR1D were analyzed for an association with syphilis in the patients and healthy people who belonged to KIR gene haplotype A. Results Of 192 healthy individuals, 187 were identified with a KIR2DS4 gene. And 91 individuals were classified as homozygous haplotype A with the percent of 48.7% (91/187) in 187 KIR2DS4 positive individuals. Of 190 syphilis patients, 181 were identified with a KIR2DS4 gene. And 89 individuals were classified as homozygous haplotype A with the percent of 49.2% (89/181) in 181 KIR2DS4 positive individuals. The frequency of KIR1D/KIR1D in syphilis patients classified as haplotype A was 16.9%, and was significantly higher than that in the control group (6.6%, P=0.032). However, there was no significant difference for the frequencies of KIR2DS4/KIR2DS4 and KIR2DS4/KIR1D between the two groups (P>0.05). Conclusion KIR1D/KIR1D might be associated with syphilis in the comparison between syphilis patients and unrelated healthy controls who were classified as homozygous haplotype A.
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Objective To develop a PCR-SSP method for detection of HLA-DRB1 alleles in the patients who were hypersensitive to Platanus Acerifolia pollen allergen,and to probe into the association between the atopic subjects to Platanus Acerifolia pollen allergen and HLA-DRB1 alleles.Methods DNA in whole blood was extracted by phenol-chloroform method.Eight pairs of specific primers for alleles were synthesized,and HLA-DRB1*0401,*0402,*0403,*0404,*0405,*0406,*0407,*0408 alleles in 20 atopic patients and 36 healthy individuals of Jiangsu Province with Han nationality were detected by PCR-SSP(polymerase chain reaction-sequence specific primer).Results By optimizing the experimental conditions PCR-SSP methods for detection of the 8 alleles were established and the distributing data of above-mentioned HLA DRB1 were obtained.The frequency of HLA DRB1*0405 and *0406 in the patients group was higher than that of in healthy controls group,while the frequency of HLA DRB1*0402 in the patients group was lower than that in controls.No significant deference for the other 5 alleles was found between the 2 groups.Conclusion HLA-DRB1*0406和*0405 seems to be the likely suspected candidate alleles responsible for susceptibility to Platanus Acerifolia pollen allergen in the atopic patients,while DRB1*0402 might be contribute to the related resistance to the allergen.
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Summary: In order to investigate the association of genotypes of HLA-DRB1 and HLA-DQB1 alleles with the genetic susceptibility of chronic urticaria (CU), genotypes of HLA-DRB1 and HLA-DQB1 genes were detected by polymerase chain reactions with sequence-specific primers (PCR-SSP) in 42 patients with CU (19 men and 23 women, mean age 30.67±12.45 y old as well as 193 racially matched healthy persons in ethnic Han from Hubei provinece. Gene frequencies of HLA-DRB1*12, *0901 (RR=3.11, χ2=7.579, P=0.006; RR=2.47, χ2=5.684, P=0.017) were significantly increased in CU patients as compared with that in healthy people. Gene frequencies of HLA-DQB1*05 (RR＝0.26, χ2=6.683, P=0.01) were significantly decreased in CU patients. It was suggested that CU was found strongly associated with HLA-DRB1*12, *0901 and HLA-DQB1*05, the former might be the genetic markers for susceptibility to CU, but the latter might play a resistive role.
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Objective:To investigate the association of human leucocyte antigen-DRB1 alleles with chronic hepatitis B and hepatic cirrhosis in Han population of Jilin area.Methods:HLA-DRB1 alleles in 61 patients with chronic hepatitis B,44 patients with hepatic cirrhosis,32 spontaneous rehabilitees after HBV infectin,and 50 healthy controls were analyzed by using polymerase chain reaction/sequence specific primer technique.Results:The frequency of HLA-DRB1*1201-3 allele in chronic hepatitis B group was higher than that in healthy control group(17.21% vs.8.00%,P=0.0427,RR=2.391).The frequency of HLA-DRB1*0701 allele in chronic hepatitis B group and hepatic cirrhosis group was markedly higher than that in healthy controls group(11.48 % vs.2.00%,P=0.006 6,RR=6.35;12.50% vs.2.00%,P=0.004 6,RR=7.00)and in spontaneous rehabilitee group (11.48% vs.1.56%,P=0.0183,RR=8.17;12.50% vs.1.56%,P=0.0136,RR=9.00).No significant differences frequencies of the other HLA-DRB1 alleles were observed among the four groups.Conclusion:Alleles of HLA-DRB1*1201-3 and HLA-DRB1*0701 are closely associated with chronic hepatitis B and maybe susceptible genes or linkage genes of chronic hepatitis B.HLA-DRB1*0701 is also closely associated with hepatic cirrhosis and maybe a susceptible gene or linkage gene-posthepatitis B-cirrhosis in Han population of Jilin area.
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BACKGROUND: To identify the human platelet antigens (HPA) associated with neonatal alloimmune thrombocytopenia (NATP), posttransfusion purpura (PTP), and platelet refractoriness, polymerase chain reaction-sequence specific primer (PCR-SSP) method and immunofluorescent method by flow cytometry were used. The frequencies of the genonotypes of HPA systems by PCR-SSP method and those of phenotypes by flow cytometry were determined. Then both types were compared each other and each types were compared with those of established reports. METHOD: Platelet suspensions were prepared from peripheral blood specimens of 200 blood donors and DNA specimens were extracted from those of 160 donors among them. Phenotypes of 200 specimens and genotypes of 160 ones were tested by flow cytometry and PCR-SSP method, respectively. RESLUTS: Frequencies of penotypes of HPA-1a, -3a, -4a, -4b and NaKa were 100.0%, 88.0, 100.0%, 0.5% and 94.0%, respectively. HPA-5 system could not be identified due to a few antigenic sites of HPA-5 systems. The genotype fequencies are of HPA-2 were a+b- 63.75%, a+b+ 35.00%, a-b+ 1.25%; HPA-3, a+b- 38.12%, a+b+ 48.13%, a-b+ 13.75%; HPA-4, a+b- 100.00%, a+b+ 0.00%, a-b+ 0.00%; HPA-5, a+b- 98.12%, a+b+ 1.88%, a-b+ 0.00%. The frequencies of HPA-4 and -5 were almost same as those of other reports but the frequencies of HPA-2 and -3 were somewhat different from others. Concordant rate between phenotype and genotype of HPA-3a,-4a and -4b were 95.6%, 100% and 99.4%, respectively. CONCLUSION: Phenotyping method by flow cytometry was rapid and objective for identifying HPA systems except HPA-5 system which has a few antigenic sites on platelet membrane. Especially it will be useful method for screening HPA-4b as possible cause of NATP in Koreans. But like other serologic methods, phenotyping by flow cytometry also require the highly qualified antiserum and appropriate amount of platelet. PCR-SSP method was also rapid and simple to test of genotypes of HPA-2~5 systems. Because PCR-SSP method is thought to be one of the most simple and economic genotyping methods to overcome the shortages of serologic methods, it is suggested to be the efficient screening method of HPA systems substituting the serologic methods in the cases which HPA sytems can not be identified by flow cytometry.
Subject(s)
Humans , Antigens, Human Platelet , Blood Donors , Blood Platelets , DNA , Flow Cytometry , Genotype , Mass Screening , Membranes , Phenotype , Purpura , Suspensions , Thrombocytopenia, Neonatal Alloimmune , Tissue DonorsABSTRACT
Objective To explore the potential association of HLA-A alleles and genetic susceptibility with systemic lupus erythematosus (SLE). Methods Polymerase chain reaction-sequence specific primer (PCR-SSP) was used to analyze the distribution of HLA-A alleles among 106 patients with systemic lupus erythematosus and 122 healthy persons. Results Nineteen out of twenty-four kinds of HLA-A alleles were found from the specimens, including 18 kinds in SLE specimens, and 15 kinds in control specimens. Among them, HLA-A*11 allele was positively associated with SLE (RR = 2.4380, EF = 0.1502, ?2 = 12.2440, P = 0.0005, Pc = 0.0095). For A*01 and A*24, although the P values were less than 0.05, the Pc values were more than 0.05 (0.9462 or 0.2356, respectively). Conclusions The results indicate that HLA-A*11 may be the susceptible allele or may be closely linked with the susceptible genes in Chinese SLE patients.
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Objective:To investigate frequencies and polymorphism of HLA-DRB1 and DQB1 allele in the Hans of Zunyi area.Methods:Polymerase chain reaction-sequence specific primers(PCR-SSP) were used to type HLA-DRB1 and DQB1 genes of 200 unrelated healthy Han individuals in Zunyi area.Results:13 HLA-DRB1 and 7 HLA-DQB1 alleles were obtained at low resolution level in all subjects.The allele DRB1*09,DRB1*08 and DQB1*05 were showed high distributing frequencies;The allele DRB1*10 and DQB1*04 were scarcely found with low distributing frequencies.Comparied with Northern and Southern Han people,it would seem that Han people in Zunyi are more closely related to the Southern ones.The allele B*07 was scarcely found in the Southern Han with a high distributing frequency(GF=2.0%).Conclusion:HLA-DRB1 and DQB1 of Han people in Zunyi have plenty of polymorphisms.They seem to distribute in line with the Southern Han's characteristics but have their own territory feature with a high B*07 frequency.