Efficient measures for blood donor recruitment, under the COVID-19 epidemic / 中国输血杂志

【Objective】 To explore ways to recruit blood donors effectively, so as to ensure blood supply for hospitals in Shenzhen with good compliance with the requirements concerning COVID-19 prevention and control. 【Methods】 From January 31 to March 31, 2020, a recruiting vanguard lead by Party members aimed at "COVID-19 control, blood supply security" was established. Otherwise, the traditional recruiting (via telephone/SMS) and on-site(on the streets) recruiting were also enhanced. The effects of the above three recruitment methods during this period, relative to the same period in 2019, were counted and analyzed. 【Results】 A total of 19 752 blood donors were recruited, and more were recruited by on-site (15 643, 79.19%) than by telephone and SMS(4 109, 20.81%), as on-site recruitment was more persuasive than the traditional one. The success rates of telephone recruitment and SMS recruitment were 11.66% (2 034 / 17 448) and 3.92% (2 075/52 915), respectively, and both were significantly higher than that in the same period in 2019(7.55% and 2.36%) (P0.05). 【Conclusion】 In times of emergency, efficient recruiting method could be achieved by grouping a vanguard, conducting scientific measures, and carrying out effect evaluation. Besides, preference could be given to donors with local registered residence, middle-age (30~50 years old) and better educated background.

Identification of a novel KIR3DL3*064 allele by cDNA cloning and sequencing / 中华医学遗传学杂志

OBJECTIVE@#To report on a novel KIR3DL3 allele identified in a southern Han Chinese individual.@*METHODS@#Peripheral blood sample was collected from a voluntary blood donor with inconclusive result by KIR3DL3 sequence-based typing (SBT). Total mRNA was extracted and subjected to reverse transcription to obtain KIR3DL3 cDNA, which was then amplified by PCR with a pair of KIR3DL3-specific primers. The product was subjected to cDNA cloning and sequencing.@*RESULTS@#cDNA cloning and sequencing have identified a wide-type KIR3DL3*00802 allele and a novel KIR3DL3*064 allele. The latter differed from KIR3DL3*00601 by a missense variant at codon 374[c.1184 C>T (p.Thr374Ile)] in exon 9. The novel KIR3DL3 allele has been officially assigned by the KIR subcommittee of World Health Organization Nomenclature Committee for factors of HLA system.@*CONCLUSION@#cDNA cloning and sequencing may be used to distinguish inconclusive results in KIR3DL3 SBT in order to identify novel KIR alleles.

Establishment of a quality control system for HLA allele typing and its key points / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To list the key points for quality control during HLA-A, B, C, DRB1 and DQB1 allele typing by taking consideration of hardware, software and experimental procedures.</p><p><b>METHODS</b>A total of 10 167 samples from randomly selected healthy blood donors and donor-recipient pairs from Shenzhen were typed for exons 2-4 of HLA-A, B, C, exon 2 of HLA-DRB1, and exons 2 and 3 of HLA-DQB1 by PCR- sequence-based typing. For 56 cases whose forward and reverse sequences were inconsistent, the samples were re-checked by a PCR-sequence specific oligonucleotide probe method. Novel alleles not included in the IMGT/HLA database were cloned and sequenced using in-house primers.</p><p><b>RESULTS</b>Eight novel HLA alleles were identified. A table for key positions of single nucleotide polymorphisms (SNPs) were generated, which summarized the key points for quality control during HLA-A, B, C, DRB1 and DQB1 allele typing. Among the listed SNPs, 3 were located at the HLA-A locus, 8 were at the HLA-B locus, 6 were at the C locus, 6 were at the DQB1 locus, and 4 were at the DRB1 locus. To ensure the quality control, an unique sample number for DNA transferring tubes in the process of experiment should be considered.</p><p><b>CONCLUSION</b>A protocol for quality control should be enforced by checking all of the key points. The SNPs and critical control points of the alleles should be examined to ensure the accuracy of HLA typing results.</p>

Establishment of a direct detection method for human platelet antigen1-17,Cab allele whole blood / 国际检验医学杂志

Objective To establish an acurate and convenient method to distinguish human platelet antigen(HPA) SNPs based on Target Enriched Multiplex-PCR(TEM-PCR),fluorescent probe melting curve analysis and blood direct PCR.Methods Design TEM-PCR primers and probes of HPA1-17,Cab alleles,amplify target sequences of all 18 alleles by blood direct PCR and distinguish different SNPs by melting curve of probes.Results The TEM-PCR could amplify all target sequences of 18 alleles and the melting curve analysis could distinguish those SNPs,the accuracy was equal to PCR-SSP method and the process was more convenient without blood genomic DNA extraction and subsequent gel electrophoresis thus decrease the cross-contamination risk.Conclusion Successfully established a HPA1-17,Cab alleles distinguishing method based on TEM-PCR,blood direct PCR and fluorescent probe melting curve analysis technique.

Analysis on the results of domestic HCC RNA genotype diagnostic kit / 国际检验医学杂志

Objective To evaluated the HCV genotyping results which obtained by genotype diagnostic kit in Shenzhen area. Methods 158 samples which ELISA test of anti-HCV were positive were collected from voluntary blood donors from 2014 to 2015,and were tested by PCR fluorescence probe method for viral load.The samples which viral load were greater than 1.0 ×103 IU/mL were then tested by HCV RNA genotype diagnostic kit.To analysis the proportion of different genotypes and the correla-tion between genotypes with vrial load.Results 54 HCV RNA reactive sample were quantity by PCR fluorescence probe method from 158 anti-HCV positive samples.The genotyping data for 45 cases which vrial load greater than 1.0×103 IU/mL were obtained by HCV RNA genotype diagnostic kit.The frequencies HCV genotype 1b,2,3 and 6 were 57.78%(26/45),6.67%(3/45),8.89%(4/45)and 26.67%(12/45),respectively.One-way ANOVA analysis showed that significant difference in viral loads was found be-tween different HCV genotype 1b and 2(F =2.861,P <0.05),and there was a significant difference in viral loads and anti-HCV S/CO by sex(P <0.05).Fisher′s exact test showed the significance difference between age and genotypes(P <0.05 ).Conclusion HCV 1b and 6 were the most predominant genotypes due to the higher viral load than the other subtypes among volunteer blood do-nors in Shenzhen,while the proportion of HCV 2,3 declined.

Sequencing analysis of a rare human leukocyte antigen, C*08:99, from a volunteer donor of hematopoietic stem cell transplantation / 中国组织工程研究

BACKGROUND:As the sequencing technology has been widely used and high-resolution confirmation of organ transplant matching has been gradualy developed, new human leukocyte antigen (HLA) aleles are emerging. However, the gene frequency of some genes cannot be calculated accurately, and there are rare reports. These genes are often ignored, and it is easy to misjudge their genotypes only according to gene frequency. OBJECTIVE:To test and analyze a rare alele, HLA-C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. METHODS: Genomic DNA was extracted automaticaly from the blood sample by using quick DNA purified kit and amplified by HLA-C locus commercial sequence-based typing kit. The purified PCR product was utilized as the DNA template in the sequencing reaction, and six direct sequencing reactions of PCR product covering exons 2, 3 and 4 in both directions were performed using commercial kit. Four direct sequencing reactions of PCR product covering exon 5 in both directions, exon 6 in forward direction and exon 7 in reverse direction were performed using in-house BigDye terminator cycle sequencing reaction kit. Sequencing reaction products purified by ethanol/sodium acetate/ ethylenediaminetetraacetic acid method were sequenced by ABI PrismTM3730 DNA Sequencer. RESULTS AND CONCLUSION:The alele assignment was analyzed with Assign-SBT 3.6+ software, and the sample HLA-C typing result was C*07:04, 08:99. Increasing the sequencing analysis at exons 5, 6 and 7 of HLA-C locus wil help to make clear the ambiguous SBT result and improve the accuracy of HLA-C typing when it is necessary, which shows important significance in clinical tissue matching. Cite this article:Wang DM, Zou HY, Nie DM, Gao SQ, Wang F.Sequencing analysis of a rare human leukocyte antigen, C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):102-106.

Studies on the Insoluble Particles in Traditional Chinese Drugs for IV Injection / 中国药房

OBJECTIVE:To observe the changes of insoluble particles in traditional Chinese drugs injections(TCDI)mixed with infusion fluid and to study the way to solve METHODS:61 kinds of TCDI in therapeutic dosages were mixed in 0 9% sodium chloride solution,then the insoluble particles formed with diameters of 2 5?5 0?10 0 and 25 0?m were counted with Coulter counter,and determined with physical method and microscope The precise filter for liquid medicine were investigated in respect to its flow rate,quantity of flow,adsorbability and scavenging action RESULTS:(1)The number of insoluble particles in 26 kinds of TCDI exceeded the standard in ChP accounting for 42 6% of total samples observed (2)The insoluble particles included glass fragments,active carbon,rubber particles,soft flocks and residue of drugs (3)The flow rate and quantity of flow met the clinical requirement with a scavenging rate of 88 5%,and no adsorbability was found CONCLUSION:Precise infilter for liquid medicine can scavenge the particles in TCDI so as to ensure the safe use of drugs for patients