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The Experience of RHD Genotyping in D-negative Blood Donors d阴性献血者RHD基因分型的经验
Pub Date : 2021-08-31 DOI: 10.17945/kjbt.2021.32.2.91
Taeeun Kim, Yunju Park, Leeseul Shin, Yu Soek Jung, M. Youn, Yeongbin Kim
Background: There have been some domestic and overseas cases of anti-D alloimmunization caused by the transfusion of serologically D-negative blood. However, it is difficult to distinguish between true D-negative and DEL variants using conventional serologic typing. Therefore, we established the RHD genotyping algorithm for the detection of DEL variants and applied this algorithm to serologic D negative donors who voluntarily consented to testing. Methods: From September 2016 to December 2020, 216 RhD negative donors who were C + and/or E+ in previous serologic typing were recruited. The screening test was PCR amplification of the RHD exons 4, 7, 10, and a promotor. Based on the results of PCR screening, true D-negative samples and RHD variants (including DEL) were discriminated. When the result was a RHD variant, exon 9 was sequenced to identify the nucleotide changes. Full sequencing was performed if no mutations were detected at exon 9. Results: Among the 216 participants, 39 cases with the C − E − c + e + phenotypes that did not meet the recruitment criteria were excluded from data analysis. Among the remaining 177 samples, 68 cases (38.4%) were RHD total deletions, 35 cases (19.8%) were RHD -CE-D hybrids, and 74 cases (41.8%) were RHD variants. Among the cases of RHD variants, 73 cases (98.6%) had c.1227G > A substitutions and were confirmed as Asian-type DEL. Conclusion: Seventy-four cases of serologic D negative donors were reclassified as RHD variants by RHD genotyping. This is believed to have contributed to the improvement of transfusion safety by lowering the risk of anti-D alloimmunization in D-negative patients. (Korean J Blood Transfus 2021;32:91-101)
背景:国内外已有因输血血清学上d阴性血引起的抗d异体免疫的病例。然而,使用传统的血清学分型很难区分真正的d阴性和DEL变异。因此,我们建立了检测DEL变异的RHD基因分型算法,并将该算法应用于自愿接受检测的血清学D阴性献血者。方法:2016年9月至2020年12月,招募216名既往血清学分型为C +和/或E+的RhD阴性献血者。筛选试验为RHD外显子4、7、10和启动子的PCR扩增。根据PCR筛选结果,区分真d阴性样本和RHD变异(包括DEL)。当结果是RHD变异时,对外显子9进行测序以确定核苷酸变化。如果外显子9未检测到突变,则进行全测序。结果:在216名参与者中,39例不符合招募标准的C−E−C + E +表型被排除在数据分析之外。其余177例中,RHD全缺失68例(38.4%),RHD -CE-D杂交35例(19.8%),RHD变异体74例(41.8%)。RHD变异中,c.1227G > A置换73例(98.6%),确认为亚洲型DEL。结论:74例血清学D阴性供者通过RHD基因分型重新分类为RHD变异。这被认为有助于通过降低d阴性患者抗d异体免疫的风险来改善输血安全性。(韩国输血杂志2021;32:91-101)
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引用次数: 1
Transfusion and Plasmapheresis in Heart and Lung Transplantation in Korea 输血和血浆置换在韩国的心脏和肺移植
Pub Date : 2021-08-31 DOI: 10.17945/kjbt.2021.32.2.129
Hyun Ji Lee, Kyung-Hwa Shin, S. Jo, Hyung-Hoi Kim
To increase the success rate of heart and lung transplantation, appropriate transfusion and desensitization treatment should be performed. In each hospital, transfusion should be performed according to the patient and hospital situation and monitored to ensure that an appropriate amount of transfusion is achieved is necessary. If HLA desensitization treatment is performed using plasmapheresis and immunosuppressants, the incidence of rejection and complications after transplantation can be reduced. For desensitization treatment that considers individual patient characteristics, close cooperation between clinical medical staff, blood bank personnel, and medical staff will be required. (Korean J Blood Transfus 2021;32:129-131)
为提高心肺移植成功率,应进行适当的输血和脱敏治疗。在每家医院,应根据病人和医院的情况进行输血,并进行监测,以确保达到适当的输血量。如果使用血浆置换和免疫抑制剂进行HLA脱敏治疗,可以减少移植后排斥反应和并发症的发生率。对于考虑患者个体特征的脱敏治疗,需要临床医务人员、血库工作人员和医务人员之间的密切合作。(韩国输血杂志2021;32:129-131)
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引用次数: 2
Therapeutic Plasma Exchange in Patients with Severe Fever with Thrombocytopenia Syndrome: A Single Institution Experience 重症发热伴血小板减少综合征患者血浆置换治疗:单一机构经验
Pub Date : 2021-08-31 DOI: 10.17945/kjbt.2021.32.2.83
Eun-Hyung Yoo, A-Jin Lee, Sang-Gyung Kim, C. Jeon
Background: Severe fever with thrombocytopenia syndrome (SFTS) is a novel infection caused by the tick-borne SFTS virus. More than 200 patients are reported every year in Korea, but there is no established treatment. In patients with SFTS, therapeutic plasma exchange (TPE) can be applied. Methods: Clinical and laboratory characteristics of patients diagnosed with SFTS who underwent TPE were analyzed. The factors that can differentiate the prognosis between the patients who recovered after TPE and those who died were analyzed. Results: Ten patients were diagnosed with SFTS and treated with TPE. The mean age was 70.8 (49–85) years, with three men and seven females. The laboratory findings showed a decrease in white blood cell (WBC) count, platelet count, and serum albumin and an increase in AST, ALT, LDH, and CK levels. Patients performed an average of three (2∼4) TPE procedures at intervals of 1∼2 days, three of whom died. Compared to the results at admission, the WBC counts increased after TPE, and the platelet counts remained unchanged. The AST, LDH, and CK levels decreased by 2∼6 fold in the recovered patients and increased in those who died. Among them, the change in LDH was statistically significant between the two groups (P=0.0227). Conclusion: TPE has been used as an adjuvant treatment in SFTS patients who do not have a definitive treatment to date. Additional studies, including small-scale studies such as this study, will be needed to establish the timing, interval, and predictive factors of the effect of TPE. (Korean J Blood Transfus 2021;32:83-90)
背景:发热伴血小板减少综合征(SFTS)是一种由蜱传SFTS病毒引起的新型感染。据报道,韩国每年有200多名患者,但没有确定的治疗方法。对于SFTS患者,可以应用治疗性血浆置换(TPE)。方法:分析经TPE治疗的SFTS患者的临床和实验室特点。分析TPE术后恢复患者与死亡患者预后的影响因素。结果:10例患者确诊为SFTS,经TPE治疗。平均年龄70.8岁(49 ~ 85岁),男3人,女7人。实验室结果显示白细胞(WBC)计数、血小板计数和血清白蛋白减少,AST、ALT、LDH和CK水平增加。患者平均每隔1 ~ 2天进行3次(2 ~ 4次)TPE手术,其中3人死亡。与入院时相比,TPE后WBC计数增加,血小板计数保持不变。康复患者的AST、LDH和CK水平下降2 ~ 6倍,死亡患者则升高。其中,两组LDH变化差异有统计学意义(P=0.0227)。结论:TPE已被用作辅助治疗的SFTS患者谁没有明确的治疗日期。需要更多的研究,包括像本研究这样的小规模研究,来确定TPE效果的时间、间隔和预测因素。(韩国输血杂志2021;32:83-90)
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引用次数: 0
Development of a Secondary Order Program for Pretransfusion Tests to Improve the Work Efficiency of Blood Bank 制定二级输血前检测方案,提高血库工作效率
Pub Date : 2021-04-30 DOI: 10.17945/KJBT.2021.32.1.22
Jung-ah Kim, J. Shin, Do Lee Lee, W. Shin, Jieun Kim, H. I. Bang
blood bank, if the Ab screening test results are positive in the pretransfusion test, an Ab identification test and polyspecific direct antiglobulin test (DAT) are performed. IgG and C3 monospecific DATs are also performed if the polyspecific DAT is positive. To perform additional tests, clinical technologists used to communicate with the clinical department by telephone, and then the clinical doctor issued the orders. There could be problems with this process, such as clerical errors and reduced work efficiency. Therefore, this study developed the secondary order program to improve the work efficiency of the blood bank. Methods: The secondary order program that allows the laboratory medicine doctors to issue additional test orders, print out barcodes in blood bank, and immediately report the results to the EMR (Electronic Medical Record) was developed. Before (Jul 2018 ∼ Jun 2019) and after (Jul 2019 ∼ Jun 2020) applying the program to inpatients, the number of telephone calls, Ab screening tests, Ab identification tests, polyspecific DAT, and monospecific DAT were compared to evaluate the effectiveness of the program. Results: After applying the program, 515 calls per year (average 43 calls per month) were reduced. Before the program, the results of 68 Ab screening tests and 16 polyspecific DATs were not reported on EMR, but no case was missed after the program. Conclusion: Through the secondary order program, the work efficiency of the blood bank was improved. It is expected that expanding this program to other blood bank tests will help implement tests faster and make them more systematic. (Korean J Blood Transfus 2021;32:22-27)
如果在输血前试验中Ab筛选试验结果为阳性,则进行Ab鉴定试验和多特异性直接抗球蛋白试验(DAT)。如果多特异性DAT阳性,也进行IgG和C3单特异性DAT检测。为了进行额外的检查,临床技术人员通常通过电话与临床部门沟通,然后由临床医生发出命令。这个过程可能会有问题,比如文书错误和工作效率降低。因此,本研究开发二级程序,以提高血库的工作效率。方法:开发二级指令程序,允许检验医学医生发出附加检测指令,在血库中打印条形码,并立即将结果报告给电子病历。在将该方案应用于住院患者之前(2018年7月~ 2019年6月)和之后(2019年7月~ 2020年6月),比较了电话次数、抗体筛选试验、抗体鉴定试验、多特异性DAT和单特异性DAT,以评估该方案的有效性。结果:应用该方案后,每年减少515个电话(平均每月43个电话)。项目实施前,有68例Ab筛选试验和16例多特异性dat未在EMR上报告结果,但项目实施后没有遗漏病例。结论:通过二级排序程序,提高了血库的工作效率。预计将这一项目扩展到其他血库测试将有助于更快地实施测试并使其更加系统化。(韩国J输血2021;32:22-27)
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引用次数: 0
Analysis of HCV Genotype with Low Titer of HCV RNA Using the Methods of Concentration 浓度法分析低滴度HCV RNA基因型
Pub Date : 2021-04-30 DOI: 10.17945/KJBT.2021.32.1.43
Jungwon Kang, J. Kang, Dae Ho Ko, M. Youn, So‐Yong Kwon
Analysis of HCV genotypes can help identify infection routes and the development of treatment methods. However, in some samples with a low titer of HCV RNA, it is difficult to analyze their genotypes. In our previous study about HCV genotyping, we could not identify 12 cases among the 175 HCV NAT reactive samples due to their low titer. In this study, we adopted three different kinds of virus concentration methods to identify the genotypes of the 12 unidentified cases and compared their efficacy. The three virus concentration methods were automatic nucleic acid extraction, polyethyleneimine-magnetic bead-based extraction, and sucrose cushion ultracentrifugation. After virus concentration using every three methods, we analyzed HCV RNA genotypes using the concentrated sample of the best efficacy. Among the 12 cases, six were identified as 1b, four as mixed types, and two were unidentified. Here we could validate that the sample concentration method is useful to identify the HCV genotypes, especially in samples with low HCV RNA titers. Furthermore, considering the convenience, high efficacy, and time-saving, automatic nucleic acid extraction is considered the most useful concentration method for samples with titer lower than 50 IU/mL. (Korean J Blood Transfus 2021;32:43-48)
分析HCV基因型有助于确定感染途径和制定治疗方法。然而,在一些HCV RNA滴度较低的样本中,很难分析其基因型。在我们之前的HCV基因分型研究中,175例HCV NAT反应样本中有12例由于滴度低而无法鉴定。本研究采用三种不同的病毒浓缩方法对12例未确诊病例进行基因型鉴定,并比较其效果。三种病毒浓缩方法分别是自动核酸提取法、聚乙烯亚胺-磁珠提取法和蔗糖垫超离心法。三种方法浓缩后,采用浓缩后效果最好的样品分析HCV RNA基因型。12例中,6例为1b型,4例为混合型,2例为不明型。在这里,我们可以验证样品浓度法对HCV基因型的鉴定是有用的,特别是在低HCV RNA滴度的样品中。此外,考虑到自动核酸提取方便、高效、省时,对于滴度低于50 IU/mL的样品,自动核酸提取被认为是最有用的浓度方法。(韩国J输血2021;32:43-48)
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引用次数: 0
Blood Coagulation Products and Other Hemostatic Agents 血液凝固产品和其他止血剂
Pub Date : 2021-04-30 DOI: 10.17945/KJBT.2021.32.1.11
Jaehyun Kim
Normal hemostasis is achieved by the concerted interactions of vascular endothelial cells, platelets, and plasma coagulation factors. The blood coagulation mechanism involves the regulated sequence of proteolytic activation of a series of blood coagulation zymogen factors. Some of these blood coagulation factors, either purified from human plasma or produced by recombinant DNA technologies, are used to treat patients with congenital or acquired deficiencies in coagulation factors. Furthermore, blood coagulation products are used for perioperative coagulation management and toical hemostatic agents to help control bleeding during surgery. Thus, the coagulant blood products have great medical importance to treat life-threatening bleeding disorders and serious medical conditions. This review provides an overview and summarizes the clinical use of coagulant blood products. (Korean J Blood Transfus 2021;32:11-21)
正常的止血是通过血管内皮细胞、血小板和血浆凝血因子的协同作用来实现的。凝血机制涉及一系列凝血酶原因子的蛋白水解激活的调控序列。其中一些凝血因子,从人血浆中纯化或通过重组DNA技术生产,用于治疗先天性或获得性凝血因子缺乏的患者。此外,凝血产品用于围手术期凝血管理和局部止血剂,以帮助控制手术期间的出血。因此,凝血产品在治疗危及生命的出血性疾病和严重疾病方面具有重要的医学意义。本文综述并总结了凝血制品的临床应用。(韩国输血杂志2021;32:11-21)
{"title":"Blood Coagulation Products and Other Hemostatic Agents","authors":"Jaehyun Kim","doi":"10.17945/KJBT.2021.32.1.11","DOIUrl":"https://doi.org/10.17945/KJBT.2021.32.1.11","url":null,"abstract":"Normal hemostasis is achieved by the concerted interactions of vascular endothelial cells, platelets, and plasma coagulation factors. The blood coagulation mechanism involves the regulated sequence of proteolytic activation of a series of blood coagulation zymogen factors. Some of these blood coagulation factors, either purified from human plasma or produced by recombinant DNA technologies, are used to treat patients with congenital or acquired deficiencies in coagulation factors. Furthermore, blood coagulation products are used for perioperative coagulation management and toical hemostatic agents to help control bleeding during surgery. Thus, the coagulant blood products have great medical importance to treat life-threatening bleeding disorders and serious medical conditions. This review provides an overview and summarizes the clinical use of coagulant blood products. (Korean J Blood Transfus 2021;32:11-21)","PeriodicalId":231122,"journal":{"name":"The Korean Journal of Blood Transfusion","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115458379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Current Status of Korean Red Cross HLA-Matched Platelet Donor Registry 韩国红十字会hla匹配血小板捐献登记的现状
Pub Date : 2021-04-30 DOI: 10.17945/KJBT.2021.32.1.1
Yeongbin Kim, A. Lim, Tae Eun Kim, Chul Ho Jung, M. Park, Jiyeong Seon, K. Youn, Anissa Lee, M. Youn
Yeongbin Kim, M.D., A Hyun Lim, M.S., Tae Eun Kim, M.S., Chul Ho Jung, M.T., Minhui Park, M.T., Jiyeong Seon, M.T., Kyoung Won Youn, M.S., An Gyo Lee, M.T., Miae Youn, M.D. Blood Transfusion Research Institute, Korean Red Cross, Wonju; Plasma Fractionation Center, Korean Red Cross, Eumseong; Chungbuk Blood Center, Korean Red Cross, Cheongju; Daejeon Sejong Chungnam Blood Center, Korean Red Cross, Daejeon, Korea
金永彬,M.D.,林爱贤,m.s.,金泰恩,m.s.,郑哲浩,m.t.,朴敏慧,m.t.,宣智英,m.t.,尹京元,m.s.,李安教,m.t.,尹美,M.D.,原州大韩红十字会输血研究所;阴城大韩红十字会血浆分离中心;清州市大韩红十字会忠北血液中心;大韩红十字会大田世宗忠南血液中心,大田
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引用次数: 1
Transfusion Dependency in Patients with Acute Myeloid Leukemia during Induction Chemotherapy 急性髓系白血病患者诱导化疗期间的输血依赖
Pub Date : 2021-04-30 DOI: 10.17945/KJBT.2021.32.1.35
H. Park, K. Shin, B. Son
Background: Blood transfusion is frequently performed as a supportive therapy during the diagnosis and chemotherapy of acute myeloid leukemia (AML). This study examined the frequency of blood transfusion and analyzed the correlation with the treatment response during induction therapy in patients with AML. Methods: From January 2018 to December 2020, blood transfusion information was collected from 23 patients diagnosed with AML during induction therapy. The frequency and volumes of blood transfusions according to the treatment response were collected and analyzed with the overall survival retrospectively. Results: The blood transfusion was performed in all patients with AML during induction therapy. The transfusion frequency and volumes were a median of five (1∼13) times and nine (2∼27) units for red blood cells, respectively. In the platelets, the median frequency was seven (2∼21) times, and the transfusion volumes were 42 (12∼128) units. At the time of the treatment response evaluation, the transfusion dependence was 0% in morphological complete remission and 20% in the morphological leukemic-free state for both RBC and platelets, and 78% for RBC and 67% for platelets in treatment failure. Although not statistically significant, transfusion independence for more than eight weeks after induction therapy showed a better overall survival (P=0.312). Conclusion: When the treatment response was good, the dependence on blood transfusion decreased. The transfusion frequency is expected to help predict the patient's treatment response and prognosis along with the peripheral blood counts. (Korean J Blood Transfus 2021;32:35-42)
背景:在急性髓性白血病(AML)的诊断和化疗过程中,输血经常作为一种支持治疗。本研究检测了急性髓系白血病患者诱导治疗期间的输血频率,并分析了输血频率与治疗反应的相关性。方法:收集2018年1月至2020年12月23例AML诱导治疗患者的输血信息。根据治疗效果收集输血次数和输血量,并与总生存率进行回顾性分析。结果:所有急性髓系白血病患者在诱导治疗期间均输血。红细胞输血次数和输血量的中位数分别为5(1 ~ 13)次和9(2 ~ 27)个单位。在血小板中,中位频率为7(2 ~ 21)次,输血量为42(12 ~ 128)单位。在治疗反应评估时,形态学完全缓解的输血依赖性为0%,形态学无白血病状态的输血依赖性为20%,治疗失败的输血依赖性为78%,血小板依赖性为67%。虽然没有统计学意义,但诱导治疗后超过8周的输血独立性显示出更好的总生存率(P=0.312)。结论:治疗效果好时,患者对输血的依赖性降低。输血频率有望与外周血计数一起帮助预测患者的治疗反应和预后。(韩国J输血2021;32:35-42)
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引用次数: 0
Case of Suspected Occult Hepatitis B Virus Infection Mistaken for a Transfusion-Transmitted Infection in a Hemodialysis Patient 一例血液透析患者疑似隐性乙型肝炎病毒感染误诊为输血传播感染
Pub Date : 2021-04-30 DOI: 10.17945/KJBT.2021.32.1.49
So Mi Kim, S. H. Kim
A 73-year-old male patient undergoing hemodialysis showed an abnormal liver function test after the transfusion of two units of red blood cell (RBC). The results of the pre-transfusion test were negative HBsAg and positive anti-HBs. On the other hand, the results of the post-transfusion test were positive HBsAg and negative anti-HBs. The other test results were positive HBeAg, positive HBV DNA, and positive anti-HBc IgG. It was reported as a blood transfusion reaction in doubt of a transfusion-transmitted hepatitis B virus (HBV) infection. An examination of the blood information management system data, health insurance data, medical records, and test of storage samples showed that it was not a blood transfusion reaction. It was believed to be an occult HBV infection in an immunosuppressive patient receiving hemodialysis. Hemodialysis patients have a high prevalence of hepatitis B and C, and are classified as a risk group of occult infections. This raises the need for HBV vaccination in dialysis patients. Virus infections can be activated in dialysis patients due to the patient's immune system and other causes. If a blood transfusion is performed during this period, attention is needed because it can be mistaken for viral transmission from blood products. (Korean J Blood Transfus 2021;32:49-54)
一位73岁的男性患者在接受血液透析后,输了两个单位的红细胞(RBC),结果显示肝功能异常。输血前检测结果为HBsAg阴性,anti-HBs阳性。另一方面,输血后检测结果为HBsAg阳性,anti-HBs阴性。其他检测结果为HBeAg阳性、HBV DNA阳性、抗hbc IgG阳性。据报道,这是一例输血反应,怀疑是输血传播的乙型肝炎病毒(HBV)感染。经检查血液信息管理系统数据、健康保险数据、医疗记录和存储样本检测,表明该事件不是输血反应。它被认为是在接受血液透析的免疫抑制患者的隐匿性HBV感染。血液透析患者乙型和丙型肝炎患病率高,被列为隐匿性感染的危险群体。这就提高了透析患者接种乙肝疫苗的必要性。由于患者的免疫系统和其他原因,病毒感染可能在透析患者中被激活。如果在此期间输血,则需要引起注意,因为它可能被误认为是血液制品的病毒传播。(韩国J输血2021;32:49-54)
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引用次数: 0
Detection of RHD 1227G>A and 1222T>C Using PCR-Restriction Fragment Length Polymorphism pcr -限制性片段长度多态性检测RHD 1227G>A和1222T>C
Pub Date : 2021-04-30 DOI: 10.17945/KJBT.2021.32.1.28
W. Kim, Geon Park
Background: DEL is an RhD variant that cannot be detected by routine serologic tests because of the extremely low expression of the RhD antigen. Detecting the common genotypes of RHD 1227G > A and 1222T > C in Korean DEL is important for safe and efficient blood transfusions. Therefore, in this study, a PCR-restriction enzyme fragment polymorphism (RFLP) method was applied to detect RHD 1227G > A and 1222T > C. Methods: DNA extracted from the blood of each segment of 56 units of RhD-negative red blood cell were used. The promoter, exon 7 and exon 9 of RHD , and exon 9 of RHCE were amplified. The PCR products of RHD exon 7, RHD exon 9, and RHCE exon 9 were treated with the restriction enzymes HpyAV and MspI, and the RFLP patterns were observed by electrophoresis. The results of PCR-RFLP of RHD exon 9 were confirmed by PCR-direct sequencing. Results: RHCE exon 9 was amplified in all 56 DNAs. RHD promoters, exon 7, and exon 9 were all amplified in 10 samples, RHD promoter, exon 7, and exon 9 were not amplified in 38 samples, and RHD promoter only was amplified in eight samples. As a result of the RHD exon 9 PCR-RFLP performed on 10 samples with all targets amplified, 10 samples were determined to be 9 samples with 1227G > A and 1 sample with 1222T > C. The PCR-RFLP result and the sequencing result were 100% identical. Conclusion: PCR-RFLP using HpyAV and MspI is a reliable and applicable method for detecting RHD 1227G>A and 1222T > C in serologically RhD negative samples. (Korean J Blood Transfus 2021;32:28-34) binding sites indicate ranges of primer binding sequences according to reference sequences (accession No. RHD NG_007494.1 and RHCE NG_009208.3). Abbreviations: Ta, annealing temperature; P, PCR; R, restriction fragment length polymorphism; S, sequencing.
背景:DEL是一种RhD变异,由于RhD抗原的极低表达,常规血清学检测无法检测到。检测韩国DEL患者RHD 1227G > A和1222T > C共同基因型对安全高效输血具有重要意义。因此,本研究采用pcr -限制性内切酶片段多态性(RFLP)方法检测RHD 1227G > a和1222T > c。方法:采用56单位RHD阴性红细胞各段血液中提取的DNA。RHD的启动子、外显子7、外显子9和RHCE的外显子9被扩增。将RHD外显子7、RHD外显子9和RHCE外显子9的PCR产物分别用HpyAV和MspI限制性内切酶处理,电泳观察RFLP模式。RHD外显子9的PCR-RFLP结果经PCR-direct测序证实。结果:56个dna中均扩增出RHCE外显子9。10份样品中RHD启动子、外显子7和外显子9均被扩增,38份样品中RHD启动子、外显子7和外显子9未被扩增,8份样品中RHD启动子仅被扩增。通过对10个样品进行RHD外显子9的PCR-RFLP扩增,10个样品确定为9个样品为1227G > a, 1个样品为1222T > c, PCR-RFLP结果与测序结果100%一致。结论:HpyAV和MspI联合PCR-RFLP检测血清学阴性RHD样品中RHD 1227G> a和1222T > C是一种可靠、适用的方法。(Korean J Blood Transfus 2021;32:28-34)结合位点根据参考序列标示引物结合序列范围。RHD NG_007494.1和RHCE NG_009208.3)。缩写:Ta,退火温度;P, PCR;R,限制性片段长度多态性;年代,测序。
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引用次数: 0
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The Korean Journal of Blood Transfusion
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