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Objective To analysis the genomic sequence of a novel human leukocyte antigen (HLA)-B*3818 allele.Methods Full length genomic sequence of an unknown HLA-B allele was cloned,followed by bi-directional sequencing and the specificity of the antigen coded by this novel allele was defined by microcytotoxicity assay.The frequency and haplotype of this novel allele was acquired by population census and parentage analysis.Results The full length genomic sequence of this novel HLA-B*3818 allele with accession number FJ561482 differs from HLA-B*380201 by two nucleotide changes in exon 4 and intron 5,respectively.One change is located at nt 660 in exon 4 where C→A alternation,which results in an amino acid substitution from Asp(GAC)to Glu(GAA)at codon 196.This alternation is a new single nucleotide polymorphism compared with all other HLA-B alleles.Another is located at genomic position 2133 in intron 5(A→C).Except for this substitution,the intron sequences of HLA-B*3818 allele are identical to those of other HLA-B*38 alleles including HLA-B*380101,B*380201 and B*3814.The serological specificity of HLA-B*3818 is B38 and the frequency of this new allele is less than 0.000 5 in Chinese Han population.The parentage analysis showed the haplotype of novel allele is A*030101-Cw*010201-B*3818-DRB1*1312-DOB1*060101.Conclusion The simultaneous mutations in exon and intron were found in the Hovel HLA-B*3818 allele,and so it can present more sequence information for studies and applications associated with HIA genes by analyzing the genomic sequences of novel HLA alleles.
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OBJECTIVE To investigate and analyze the kinetics of serological marks and virus load of the follow-up blood donors. METHODS The quantitation of HBV DNA of 6 HBsAg negative and DNA positive blood donors was detected by Roche PCR Monitor,and the donors were genotyped and traced by serological tests. RESULTS(Three of 6 follow-up) donor samples were seroconverted after more than 40 days follow-up study.The viral loads were with range of(35-1.8)?10~4 copies/ml,1 of 6 donors had acute infection,the peak load was(1.8?10~4 copies/ml),3 of 6 were with lower level HBV carrier with fluctuating viral load ranged 100-500 copies/ml.One donor had decreasing viral load,when HBsAg converted to positive,virus wasn′t detected.The last one had fluctuating virus load below(100 copies/ml),intermittently falling below the threshold of the assay. CONCLUSIONS It is necessary to implement HBV DNA detection for blood screening,and further strengthen the safety of blood transfusion.
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Objective To establish the entirely automatic method of nucleic acid amplification testing (NAT) for blood screening and to study the feasibility of NAT.Methods Using entirely automated extraction method to extract nucleic acid , amplified and detected by Roche COBAS AMPLICOR system,evaluating the sensitivity and efficacy.Results The 95% limits of HBV DNA, HCV RNA/HIV-1 RNA tests by automation system were 38.9,16.4IU/ml and 20.4 copies/ml,95% Confidence Intervals were [21,323], [10.5,342] and [12,300] respectively.8 of 16 512 donations were PCR for HBV DNA positive,the DNA positive rate was 0.048%.7/8 donations were Anti-HBc positive,The last one was also converted positive.No positive HCV RNA and HIV RNA was detected. 3/6 following up samples seroconverted.Conclusions The entirely automatic system can be applied in blood screening.
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0.05).However,during the thawing process of the APCs,four out of ten and three out of eleven APCs samples appeared aggregative reaction after cryo-preservance for 4 and 5 years,respectively.The majority of the samples extensively lost their factor Ⅲ activities in the fifth year storage.When fresh,liquid platelets were stored at room temperature(22 ℃),the in vitro adhesion and agglutination gradually decreased and statistic differences were observed in 72 h(P