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The influence of specimen storage time to platelet count in unpaid blood donors of platelets

Ziyang FENG ; Lishuang ZHU ; Heshan TANG ; Baohua QIAN

Chinese Journal of Blood Transfusion.2017;30(7):704-705. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.017

Objective To analyse the effect of specimen storage time on platelet count in apheresis Donors,in order to choose the appropriate testing time.Methods We choose fifty healthy Unpaid Blood Donors of Platelets to test platelet count in 0,0.5,1,3,6h respectively by the blood counting instrument.Results At room temperature,the count of platelets from blood samples is relatively lower at 0 hour and the difference is significant (P ＜ 0.05).After 0.5-6 hours,the count of platelets become stabilized and not has significantly different(P＞0.05).Conclusion The count of platelets of blood samples is lower at 0 hour than 0.5-6 hours,this work suggest that the count of platelets of blood samples should be done at 0.5-6 h in order to protect platelets quality.

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The study on nucleic acid testing for bacterial detection and its preliminary application on concentrated platelet

Yuning LIU ; Xun WANG ; Danna FEI ; Xuzan HAO ; Shaou FU ; Yao JIA

Chinese Journal of Blood Transfusion.2017;30(7):702-704. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.016

Objective To establish a nucleic acid testing assay for bacterial detection with meeting automatic requirement,and use it preliminarily to detect bacterial contamination in concentrated platelet.Methods Optimizing the bacterial DNA extraction methods,evaluating the sensitivity and specificity of the optimized assay with different bacterial strains,and applying the assay on bacterial detection for 100 units of concentrated platelet.Results The optimized extraction methods met the criteria of automatic testing.The sensitivity of the assay is 10,150,65,15 CFU/mL for Escheriehia coli,Stphylococcus aureus,Bacillus subtilis,and Pseudomonas aeruginas respectively.There was no non-specific amplification detected.And the assay could detect 1 contaminated bacterial from 100 units of concentrated platelet as same as BactALT bacterial culture.Conclusions Nucleic acid testing for bacterial genome 16S DNA could detect common contaminated bacterial sensitively and fast from platelet product.This assay could have the same effect when compared to bacterial culture methods when it was applied in concentrated platelet products.

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Identification of anti-C alloantibodies accompanying anti-e autoantibodies

Langan PI ; Zhangyong HE

Chinese Journal of Blood Transfusion.2017;30(7):693-695. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.013

Objective To confirm the specificity of antibodies recently found in a hospitalized patient.Methods Methods including antibody screening,antibodies identification,alloantibody titers,isolation of red blood cells by high speed capillary centrifugation and determination of Rh phenotype were employed to investigate the characteristics of antibodies.Results Anti-C alloantibody and anti-e autoantibodies were detected in the patient.Autoantibodies were non-specific in the early stages,and gradually changed into specific auti-e antibodies.The titers of anti-C alloantibody were always lower than 4.Conclusion The identification of antibodies should be integrated as far as possible in various aspects of methodology.

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Establishment of reference range for thromboelastography in pregnancy

Hao TANG ; Yunfeng FU ; Rong GUI

Chinese Journal of Blood Transfusion.2017;30(7):687-689. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.011

Objective To establish a reference range for the normal value of thromboelastography (TEG) in pregnant females.Methods According to the results of pregnancy and physical examination,166 pregnant females and 64 healthy females without pregnancy were selected as the pregnant group and the non-pregnant control group,respectively.The TEG value and the traditional coagulation index were measured.The TEG parameters of the two groups were compared and analyzed,establishing a reference range for the parameters.We further analyzed the effect of full-term pregnancy on TEG results and the correlation between traditional coagulation index and TEG test results.Results The traditional coagulation index and TEG test results of the pregnant females andthe non-pregnant females were significantly different.According to the results,a new TEG reference range was established:R 3.9-7.5 min,K 1.0-2.4 min,α 57.6°-74.9°,MA 55.7-75.7 mm,LY30 0-0.56％,CI(-0.97)-3.6.Full-term pregnancy had no significant effect on TEG results.In addition to LY30,other parameters of TEG had some correlation with the traditional coagulation index.Conclusions The general TEG reference range does not apply to pregnant females and established TEG normal reference range for pregnant females can be applied for clinical use.

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Genotype analysis of ABO blood group system in 178 Chinese population of four nationalities

Yunqing YAN ; Zhiliang WANG ; Hengshi XU

Chinese Journal of Blood Transfusion.2017;30(7):684-686. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.010

Objective To establish a method for accurate typing of ABO in blood donors and patients with difficult blood types,and to provide clinical reference for safe blood transfusion,organ transplantation,and bone marrow matching.Methods The results of serological and genotyping of 178 volunteers from four ethnic groups in China were analyzed statistically,and the blood type difference coincidence test was carried out.Results The difference of blood type difference between the four nationalities was checked (x2 =24.5,P＞0.05).The results showed that there was no difference in blood group distribution between the four nationalities.Conclusion Serological results of 178 volunteers from four ethnic groups in China are consistent with the results of molecular biology,and there is no difference in blood type distribution.

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Analysis of serological and genetic test results of rare Ax14 subtypes in a young child

Xin LIU ; Jing ZHOU ; Xiuyun XU ; Xiaoyan TONG ; Fang WANG ; Liping AI ; Jin SHU ; Lina HE ; Ziheng XU ; Beini WAN ; Genshui ZHOU

Chinese Journal of Blood Transfusion.2017;30(7):682-684. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.009

Objectives To identify ABO blood type from a child having discrepant results in forward and reverse ABO blood grouping by serological identification and genetic testing.Methods After routine serological detection with ABO blood group,the ABO gene and ABO blood group-A subgroup genotype were tested by PCR-SSP method.Results The Serological results showed that the specimen was positive type A (anti-A:1 +w),the reverse type is O type (Ac:2 +,Bc:4 +);PCRSSP A-subtype typing showed that the genotype of the child was Ax14/O2.Conclusion Difficult blood type identification sometimes need to combine serology and molecular biology methods to confirm.In this case,the phenotype of the child was Ax,and the genotype was Ax14/O2.

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Molecularmechanism of Bel subtype caused by mutation p.R168W of glycosyltransferase B

Songsong GONG ; Wei ZOU ; Fang LI ; Yuqin WANG ; Hang LEI ; Xuefeng WANG ; Xiaohong CAI

Chinese Journal of Blood Transfusion.2017;30(7):679-681. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.008

Objective To study the molecular mechanism of Bel subtype caused by mutation p.R168W of glycosyltransferase B.Methods Serological test,SSP-PCR and direct sequence the Exon6 and Exon 7 of the ABO gene.Construct a 3D molecular model and predict the structural impact of GTB protein mutations.Results A antigen or B antigen can't be detected on the surface of the propositus' RBC,and only anti-A antibodies were detected in her serum.But serological test indicated her daughter's blood type was a normal B type.SSP-PCR test indicated propositus' ABO gene type is O1 B.By gene sequencing the Exon 6 and Exon 7 of the ABO gene,a ABO Bel allel(c.502C＞T,p.R168W)was discoverd in both the propositus and her daughter.Through the propositus' daughter coexisted Bel gene with normal B gene,her blood type was a normal B type.Conclusions ABO gene c.502C＞T mutations cause Bel phenotypes in patients by reducing the stability of GTB.

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A novel variation of B allele is responsible for ABweak phonetype

Jing WAGN ; Guoling YOU ; Xingchen YE ; Ping GU ; Qiuhu PAN

Chinese Journal of Blood Transfusion.2017;30(7):676-678. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.007

Objective To investigate the molecular basis of ABO gene in a patient with serologic ABO blood group discrepancy.Methods Serologic blood group identification,Coombs' test and antibody screening were detected with DG Gel Confirm cards,Neutral cards,Coombs cards by WADiana/8XT Compact Analyzer (from Diagnostic Grifols,S.A).The enhancer,promoter,exon 1 ～ 7 and their adjacent intron region of ABO gene were amplified by using polymerase chain reaction (PCR) method.Results The patient's red blood cells was determined as weak B phenotype showing two groups in gel and mixed field in tube with monoclonal anti-B,and A phenotype with monoclonal anti-A.DNA sequencing showed nine variants in ABO gene.One heterozygous variation in exon 6 (297A＞G) and eight heterozygous variations in exon 7 (467C＞T,526C ＞G,657C＞T,703G＞A,796C＞A,803G＞C,829G＞T 930G＞A) were identified and 829G＞T was the novel variant.Compared with Blood Group Antigen Gene Mutation Database,genotype of the patient was weak expression of A102/B101.Conclusion The novel variation of B allele is the main reason of Bweak phonetype in A102/B101 genotype.Serological and molecular biological detection help to understand the characteristics of blood group phenotype and genotype,provide the guidance for clinical transfusion strategies.

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Relationship between HLA antibodies produceand the number of gravidity

Feng DAI ; Rui WANG ; Mengsi HU ; Zhiwei LIU

Chinese Journal of Blood Transfusion.2017;30(7):674-676. doi:10.13303/j.cjbt.issn.1004-549x.2017.07.006

Objective Study the relationship between HLA antibodies production and the number of Gravidity.Methods A total of 856 pregnant women of first pretanal in Xiasha Hospital of Sir Run Run Shaw Hospital of Zhejiang University from Nov 2016 to Mar 2017 were recruited.The blood samples were collected during 24-28 weeks of pregnancy,and the expression of platelet-specific alloantibody was detected by solid-phase red cell adherence assay.Results There were 67 positive HLA antibody in pregnant women,and the antibody positive rate was 7.8％.There were 15 positive HLA antibody in 334 first pregnant women,14 in 214 two or more pregnancy pregnant women without abortion,10 in 107 pregnant women with once abortion,28 in 223 pregnant women with two or more abortion.The antibody positive rate respectively is 4.5％,6.5％,9.3％ and 12.6％.The production of HLA antibody was significantly associated with the numberof gravidity.Conclusion The antibody positive rate of two or more pregnancy is significantly higher than the fisrt pregnancy.

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Nucleotide Sequences Analysis of Proviral Envelope Region of Shantou Strain of Human T lymphotropic Virus Type Ⅰ in China.

Wen ZHUANG ; Zebin XIAO ; Feng CAO

Chinese Journal of Blood Transfusion.1988;0(01):-.

Objective To understand the gene characteristics of envelope region of human T lymphotropic virus type Ⅰ(HTLV Ⅰ)from Shantou region of China. Methods The gene of proviral envelope region of HTLV Ⅰ Shantou strain (named HTLV Ⅰ WHP)was amplified by PCR,using 4 designed primers,and then was sequenced,meanwhile it was compared with that of various HTLV Ⅰ representative strains from all around the world,and subsequently it was analyzed to construct its phylogenetic tree.Results The nucleotide sequence of complete envelope region of HTLV Ⅰ WHP provirus we have obtained was 1466 bp.There no deletion and insertion in the deducted amino acid sequence.The results of phylogenetic tree analysis showed that the HTLV Ⅰ WHP strain was most closed to some of the strains from Japan and Caribbean,and like the latter,belonged to the subtypea of cosmopolitan group.Conclusion The HTLV Ⅰ strains either from the coastal region of Southeast China or from Japan,Caribbean and so on have the common origin of evolution.

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