Objective:A comparison study of polymerase chain reaction (PCR) and serological typing method was used to detect HLA B27.Methods:78 samples from patients suspected with ankylosing spondylitis (AS) were genotyped by PCR and some of them were assayed by serological typing.Results:24/36 samples with HLA B27 gene were compared with serological typing ,the results of 5 samples serological typing were dubious or contradictory with PCR genotyping.21/42 samples without HLA B27 gene were compared with serological typing,the results of 3 samples serological typing were dubious.Conclusion:The method of B27 genotyping by PCR is more rapid,inexpensive and exact than phenotyping by serology.
To study the method for Rhesus box test and its significance, the sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed according to RhD gene sequence; the upstream, downstream and hybrid Rhesus boxes were determined by PCP-SSP and mismatched PCR. The results showed that this method was confirmed by DNA Standard test. It was shown that in unrelative RhD positive individuals RHD+/RHD-, RHD+/RHD+ genotype accounted for 9.00%, 91.00% respectively, and in RhD negative individuls RHD+/RHD-, RHD+/RHD+, RHD-/RHD-genotype were 26.14%, 3.92%, 69.94% respectively. It is concluded that the method of Rhesus box test was confirned to be reliable and can be used for the identification of RhD haplotype gene structure, as well as for study on inheritance, clinical transfusion and neonatal hemolytic diseases.
Objective:To compare the result of HLA A,B serological typing with DNA typing in 136 samples Methods:All the HLA A,B frequent alleles were typed by PCR SSP with 38 pairs of primers Results:60 of 136 samples(44 1%) showed discrepancies between serological and DNA typing;The error rate of HLA A serological typing was 23 3% in the homozygotes and 9 4% in the heterozygotes(total 11 2%) and at the HLA B was 47 4% and 18 4%(total 20 6%);False positive,false negative and misassignment of the HLA A serological typing were 6 6%,2 5% and 2 1% respectively,of the HLA B were 6 7%,3 6% and 10 3% respectively Conclusion:Error rate of HLA A serological typing in the homozygotes was significantly higher than in the heterozygotes(P0 050) ;Error rate of HLA B serological typing in the homozygotes was also higher than in the heterozygotes(P0 005);Error rate of HLA B serological typing was higher than that of HLA A (P0 005)
Objective:sHLA Ⅰ antigens may predict the onset of rejection after organ transplantation and monitor some diseases. The method of quantification of sHLA Ⅰ antigens and its normal level in Chinese were reported. Methods:Sandwich ELISA assay was used. After microtiter plate coated by W6/32 mAb, serum was added, followed by incubation with anti β 2m HRP and the color development. Standard curve was obtained by quantification of sHLA Ⅰstandard reagent in series dilution. The amount of sHLA Ⅰantigens was examined in 113 normal healthy volunteers. Results: sHLA Ⅰ antigens can be exactly quantificated by sandwich ELISA assay. The level of sHLA Ⅰ antigens in normal individual is 789.27±506.32 ng/ml. Conclusion: Sandwich ELISA assay is special, sensitive and reproducible for sHLA Ⅰ antigens, and the results can be compared each other only when reagents and protocols are the same.
Objective:Studied on Rh blood group in Han nationality comparison with in Uigrus by blood group and serology method, and its significance was discussed.Methods:Rh antigens were detected with monoclonal and multiclonal D,C,c,E,e antibodies routinely. Negative results with routine serology were confirmed with IAT and Del types were detected with absorption/elution test.Results:The phenotypes out of 228 cases of unrelative Hans with RhD(-) were 130 ccee(57.02%),68 Ccee(29.82%) and 11 CCee(4.82%),without CCEe and CcEE phenotypes.The ccee(88.89%),Ccee(5.56%) and CCee(5.56%) phenotypes could be detected only out of 72 cases of Uigurs with RhD (-) in Xinjiang, China, and the ratio of Uigurs in ccee and Ccee phenotypes was higher than that of Hans(P0.005), but the ratio of CCee didn't posses statistical discrepancy(P0.05),compared to that of Hans.42 cases of the Del types(18.42%) were detected out of 228 unrelative Hans with RhD(-),however, none of the Del types was obtained out of 72 unrelative Uigurs. 6 cases of weak D belonged to RhD positive with weak agglutination,and 3 cases of Dtypes were classified to weak D,but the other Dtype showed negative with saline agglutination test and weak positive with IAT(belong to low graded weak D).Conclusion: Rh bood group of Uigurs on Xinjiang posses Oriental and Caucasian characteristics,which is a special Chinese nationality.
Objective:To observe RhD gene in Chinese.Methods:RhD gene among 32 cases of serological RhD positive and 44 cases serological RhD negative were tested by PCR-SSP.Results:Exons of RhD positive subject were intact.Exons of RhD negative subject showed total deletion(61.4%),partly deletion(18.2%) and intact(20.1%).Conclusion:RhD gene exons are morphisms in Chinese.
Objective:To study enzymatical digestion of human B-like antigen of cynomolgus monkey for providing basis of preparation of human type O red cells.Methods:The B-like blood group antigen of cynomlgus monkey was digested by recombined α-galactosidase and various biochemistry parameters were tested for pre- and post-digestion red cells and it was transfused to animal.Results:The shape and various biochemistry parameters as well as function of post-digestion were normal.The blood pressure,pulse and respiration of recipient monkeys had not distinctly charged post-transfusion compared with pre-transfusion.Viabilities within 24 h of digestion cells in vivo were 96.5% and 95.0% versus 97.1% in control,T1/2 were all 16 d versus 16 d in control.Components and concentration in recipient blood and urine did not show a significant change compared with control(P0.05).Conclusion:Human B-like antigen on cynomolgus monkey could be α-galactosidase digested and it did not change the shape and function.There was no transfusion after human A-like cynomolgus monkeys received enzye-digestion huan B-like red cells,our results suggest that it may be possible to modify human ABO blood group.
Objective:To study on significance of ABO genotyping.Methods:To use the methods of polymerase chain reaction sequence specific primers(PCR SSP) for genotyping of ABO blood group and observation ABO gene polymorphism as well as identification of doubt sample.Results:The reliability of the genotyping for ABO blood group was proved by testing DNA samples previously known genotypes.The results of genotyping for 104 healthy and unrelated HAN individuals were correspond with serological phenotypes.The method of ABO genotyping was applied for clinical pre transfusion ABO typing,pre delivery fetal ABO typing,parenting test and subgrouptyping.Conclusion:The method of ABO genotyping may correct typing for doubt sample of ABO serological typing.
