ABSTRACT
Background@#The Korean Red Cross has conducted serologic tests for C, c, E, e antigens and found 18 D-- donors.In this study, we performed RHCE genotyping to identify the molecular characteristics of the serologic D-- blood type in Korean blood donors. @*Methods@#We performed RHCE-specific PCR-based electrophoresis to check the amplification pattern of each exon.Sanger sequencing was conducted to find the variants in the nucleotide sequence. We determined the RHCE genotype based on the electrophoresis and Sanger sequencing results. @*Results@#Total eight out of 18 D-- donors were participated in this research. In the PCR-based electrophoresis tests, RHCE exons 3, 4, and 6 were not amplified in samples #4, #6, and #8. Also, sample #2 showed an abnormal band pattern of RHCE exon 9. The Sanger sequencing results showed that the nucleotide sequences of the RHCE exons 5, 7, and 8 in samples #4, #6 and #8 corresponded to the nucleotide sequences of RHD exons 5, 7, and 8, respectively, suggesting the possibility of a RHCE-RHD(3-8)-RHCE hybrid allele. The nucleotide sequences of RHCE exons 7 and 8 in sample #2 were the same as the nucleotide sequences of RHD exons 7 and 8, respectively.In samples #1, #3, #5, and #7, no specific variants known to cause D-- phenotype were found. @*Conclusion@#RHCE genes partially replaced by the RHD genes were found in four out of eight participants and three of them were identified as ?RHCE*02N.07, which is known as the RHCE null allele. A further study with complete RHCE sequencing could be helpful for an understanding of the molecular mechanisms of samples in which no significant variants were identified.
ABSTRACT
HLA-matched platelet transfusion is required for patients with platelet refractoriness due to HLA alloimmunity. From 2013 to 2019, the Korean Red Cross has recruited 4,080 donors for HLA-matched platelets. The patient’s HLA information should be submitted to the Korean Red Cross in accordance with the WHO HLA serologic specificities. When HLA-matched platelets are requested, the Korean Red Cross selects the appropriate donors based on Duquesnoy’s matching grade classification (1977) and CREGs defined by Takemoto, Fuller, and Rodey (2007) and then contacts them to request blood donations. Platelets of HLA-matched donors are collected by apheresis and supplied to the hospital. To make this process more efficient, the Korean Red Cross introduced a systemic standard work procedure using a computer program for blood donor management and HLA matching. Owing to the extensive polymorphism of the HLA types, expansion of the donor pool would be required to supply HLA-matched platelets sufficiently. As the number of registered donors for HLA-matched platelets is limited, it should only be ordered when the indication criteria for its use are met. The Korean Red Cross is planning to study genotype-based matching strategies for patients with rare HLA types and receive patients’ laboratory test results from medical institutions to evaluate the effectiveness of HLA-matched platelet transfusions.
ABSTRACT
HLA-matched platelet transfusion is required for patients with platelet refractoriness due to HLA alloimmunity. From 2013 to 2019, the Korean Red Cross has recruited 4,080 donors for HLA-matched platelets. The patient’s HLA information should be submitted to the Korean Red Cross in accordance with the WHO HLA serologic specificities. When HLA-matched platelets are requested, the Korean Red Cross selects the appropriate donors based on Duquesnoy’s matching grade classification (1977) and CREGs defined by Takemoto, Fuller, and Rodey (2007) and then contacts them to request blood donations. Platelets of HLA-matched donors are collected by apheresis and supplied to the hospital. To make this process more efficient, the Korean Red Cross introduced a systemic standard work procedure using a computer program for blood donor management and HLA matching. Owing to the extensive polymorphism of the HLA types, expansion of the donor pool would be required to supply HLA-matched platelets sufficiently. As the number of registered donors for HLA-matched platelets is limited, it should only be ordered when the indication criteria for its use are met. The Korean Red Cross is planning to study genotype-based matching strategies for patients with rare HLA types and receive patients’ laboratory test results from medical institutions to evaluate the effectiveness of HLA-matched platelet transfusions.
ABSTRACT
BACKGROUND: The use of a functionally closed system for the glycerolization and deglycerolization of red blood cells (RBCs) allows for prolonged post-thaw storage for more than 24 hours. The aim of this study was to assess glycerolization and deglycerolization processing for RBCs using a high glycerol method in the automated, closed system provided by Haemonetics ACP 215. METHODS: Thirty-five packed RBCs were glycerolized using the ACP 215 to a final concentration of 40% (wt/vol). The units were either frozen as such (n=30) or excess glycerol was removed (n=5) before freezing. After storage at −80℃, the units were thawed, deglycerolized and resuspended in SAG-M. The frozen-thawed RBCs were stored at 4℃, and analyzed for their stability and in vitro quality. RESULTS: No prefreeze excess glycerol removal units showed significantly less potassium leakage during post-thaw storage compared to the prefreeze excess glycerol removal units. All measurements of the stability and in vitro quality of thawed RBCs prepared from frozen RBCs without the prefreeze removal of excess glycerol during post-thaw storage at 4℃ for 7 days were acceptable to the American Blood Bank Association's standards and European standards. CONCLUSION: RBCs frozen without prefreeze removal of excess glycerol and the ACP 215 simplifies cryopreservation procedure and increases the stability of frozen-thawed RBCs. This increases the practical applicability of cryopreserved RBCs in blood transfusion practice.
Subject(s)
Blood Banks , Blood Transfusion , Cryopreservation , Erythrocytes , Freezing , Glycerol , In Vitro Techniques , Methods , PotassiumABSTRACT
BACKGROUND: National reference standards are essential to the quality assessment and regulatory approval of in vitro diagnostic medical devices. However, the long-term stability of national reference standards has not been comprehensively secured. This study was performed to assessment on the long-term stability of the hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus-1 (HIV-1) national reference standards intended to be used for the nucleic acid amplification test (NAT). METHODS: The viral loads of the MFDS (Korea Ministry of Food and Drug Safety) working standard and recombinant DNA for HBV, HCV, and HIV-1 were measured before and after storage at −70℃ for up to 72 months using Cobas Ampliprep/Cobas Taqman assays (Roche Molecular System, Inc., Branchburg, USA) at defined time points. RESULTS: The viral loads of national reference standards for in vitro diagnostic medical devices of HBV, HCV, and HIV-1 stored at −70℃ for up to 72 months did not differ significantly from the baseline viral load. The changes in viral load of national reference standards of HBV, HCV, and HIV-1 tested after storage at −70℃ for up to 72 months ranged from −0.36 to 0.16 log10 IU/mL and did not exceed 0.5 log10, which is the estimated intra-assay variation of molecular tests. CONCLUSION: The HBV, HCV, and HIV-1 national reference standards for in vitro diagnostic medical devices intended to be used for the NAT were relatively stable after long-term storage at −70℃ for up to 72 months, regardless of the initial titer.
Subject(s)
Humans , DNA, Recombinant , Hepacivirus , Hepatitis B virus , HIV-1 , In Vitro Techniques , Nucleic Acid Amplification Techniques , Reagent Kits, Diagnostic , Viral LoadABSTRACT
The blood supply can become disrupted in situations of increased demand during unexpected national catastrophes and when a patient needs a rare blood transfusion, which depends on the blood inventory in peacetime. Cryopreservation of blood, which can be stored up to 10 years, represents a possible solution to this problem by avoiding storage lesions. This review describes frozen red cell technologies, quality control issues related to post-thaw red blood cells, and preconditions and practical considerations for implementation of a frozen blood banking system in Korea.
Subject(s)
Humans , Blood Banks , Blood Transfusion , Cryopreservation , Erythrocytes , Korea , Quality Control , Strategic StockpileABSTRACT
BACKGROUND: Transfusion transmissible emerging infectious diseases (EIDs) is a potential risk to the safety of blood transfusions due to the lack of donor screening assays. To prevent the spread of EIDs through blood transfusions, we attempted to predict the possibility of blood donations from people with EIDs using a public database. METHODS: We used the Disease Web Statistics System of the Korean Centers for Disease Control and Prevention and Korean Statistical Information Service. We estimated the possibility of blood donations from people with EIDs using the public database combined with the database made available by the Blood Information Management System of the Korean Red Cross. RESULTS: Among the transfusion transmissible EIDs, Babesiosis, Leishmaniasis, West Nile fever, Chikungunya, and Dengue fever were reported in Korea. All of them were cases imported from abroad. Although the number of reported cases of Babesiosis, Leishmaniasis, West Nile fever, and Chikungunya were less than 10 per year until 2016, the reported cases of Dengue fever gradually increased from 2001, and there were 318 cases of Dengue fever in 2016. CONCLUSION: The possibility of blood donation from people with transfusion-transmissible EIDs was low because all reported transfusion-transmissible EIDs in Korea were from foreigners and blood donation from Koreans who returned from abroad was restricted for a period of a month. Nonetheless, preventive strategy for donation from people is necessary given the recent increase in Dengue fever.
Subject(s)
Animals , Humans , Babesiosis , Blood Donors , Blood Transfusion , Communicable Diseases, Emerging , Dengue , Disease Outbreaks , Donor Selection , Emigrants and Immigrants , Information Management , Information Services , Korea , Leishmaniasis , Red Cross , West Nile FeverABSTRACT
BACKGROUND: Since 2001, the Korean Red Cross has performed malaria antibody test for blood donors in malaria-risk areas to prevent transfusion-transmitted malaria. However, due to insufficient sensitivity and specificity the malaria antibody assay is not considered an efficient screening method. Therefore, we have considered discontinuing malaria antibody testing for blood donors. METHODS: We analyzed the results of malaria antibody test from 2001 to 2014 utilizing data from the Blood Information Management System of the Korean Red Cross. RESULTS: Among 16,650,812 donations tested from 2001 to 2014, 50,143 donations (0.30%) showed positive results. However, there was no truly infected case at the time of donation. The positive rate among blood donations was between 34 and 39 per 10,000 in 2001, but between 9 and 10 per 10,000 in 2014. There was no interregional disparity in the positive rate of blood donations. CONCLUSION: Korea is in a malaria elimination phase and malaria antibody testing in limited areas is not effective, therefore we propose discontinuing the malaria antibody test.
Subject(s)
Humans , Blood Donors , Information Management , Korea , Malaria , Mass Screening , Red Cross , Sensitivity and SpecificityABSTRACT
BACKGROUND: Currently, serological assay, immunoblotting, and nucleic acid amplification test (NAT) are required as reentry tests for deferred donors with hepatitis C virus (HCV) or human immunodeficiency virus (HIV) screening reactive result. However, immunoblotting must be performed even for serological nonreactive donors. In this study, the efficacy of immunoblot applications for serological nonreactive donors in donor reentry procedures was examined. METHODS: We analyzed non-qualified donors with immunoblot results from 2011 to 2015 in Korea and investigated reentry procedures related with HCV or HIV in other countries. RESULTS: Percentages of donors who could not be released due to immunoblot results even with serological nonreactive results were 54.2% (1,824/3,367) for HCV and 35.9% (4,300/11,964). In the case of 662 donors, their results were considered to be different using other assay kits or based on other criteria. In other countries, immunoblotting is not required as a donor reentry test. CONCLUSION: Indeterminate or reactive immunoblotting results in serological nonreactive donors were due to nonspecific reactions. It is not reasonable to apply immunoblotting to serological nonreactive donors. Therefore, we suggest that immunoblot assays be excluded from the reentry test.
Subject(s)
Humans , Hepacivirus , HIV , Immunoblotting , Korea , Mass Screening , Nucleic Acid Amplification Techniques , Tissue DonorsABSTRACT
BACKGROUND: In the Korean Red Cross, anti-HTLV (Human T-cell lymphotropic virus)-1/2 screening assay has been performed in all donated blood except plasmapheresis since April 2009. For anti-HTLV-1/2 positive donors, both Western blot (WB) and nucleic acid amplification test (NAT) are performed as confirmatory assays. In this study, we evalutated the efficiency of the current confirmatory assay scheme to improve the confirmatory assay scheme for anti-HTLV1/2. METHODS: The results of the HTLV confirmatory assay from April 15th 2009 to April 14th 2015 were analyzed using the Blood Information Management System of the Korean Red Cross. We also investigated the current situation in other countries. RESULTS: Of 12,923,854 donations, 3,483 (0.027%) showed positive results in anti-HTLV-1/2. Of the 3,483 donations, 499 (14.3%) showed positive results in WB or NAT or both. The number of positive cases in both was 461. Therefore, the concordance rate was 92.4%. In the cases of positive results only in NAT, the WB results were all indeterminate (ID). Most countries are using immunoblot assay as a confirmatory assay for anti-HTLV positive blood donors. In the results, there were no cases of positive result in only NAT with a negative result in immunoblot assay. CONCLUSION: It was considered that the accomplishment of only WB as a confirmatory assay for anti-HTLV-1/2 positive donors may be sufficient in the aspect of safety and economics. However, in the case of WB ID result, it may be better to perform NAT as a supplemental test.
Subject(s)
Humans , Blood Donors , Blotting, Western , Information Management , Mass Screening , Nucleic Acid Amplification Techniques , Plasmapheresis , Red Cross , T-Lymphocytes , Tissue DonorsABSTRACT
BACKGROUND: A range of well characterized materials are needed for validating the performance of hepatitis B surface antigen (HBsAg) immunoassays. These materials are purchased currently from overseas manufacturers at a high cost and with limited quantity. This study was conducted to establish an HBsAg low titer performance panel for use as a national standard for validation of HBsAg immunoassays in Korea. METHODS: 476 plasma units reactive on blood donor screening were collected HBsAg was tested using 3 enzyme immunoassays (EIA) and 1 chemiluminescence immunoassay (CIA). Units reactive on the CIA assay or on 2 or more immunoassays were subjected to hepatitis B virus (HBV) DNA quantification, HBV genotyping and subtyping. Units reactive on HBV DNA quantification were confirmed for HBsAg by neutralization. Candidates for the panel were subjected to a collaborative study performed at 7 laboratories using 7 immunoassays. RESULTS: Eleven HBsAg positive units were selected for the low titer performance panel based on HBsAg immunoassay, HBV DNA quantification, HBV genotyping and subtyping results. The range of the HBsAg concentration of the panel members was 0.05~1.28 IU/mL. Two HBsAg negative units were also included as negative controls. CONCLUSION: As a result of this study, a low titer performance panel [KFDA standard (08/028); HBsAg low titer performance panel (BTRL HBV/LP)] for validation of HBsAg immunoassays has been established as a Korean national standard. Use of this panel will improve performance assessment of HBsAg immunoassays. Because the performance of immunoassays cannot be assessed properly with a limited number of panels, continuous efforts are needed to develop a range of performance panels.
Subject(s)
Humans , Blood Donors , DNA , Hepatitis , Hepatitis B , Hepatitis B Surface Antigens , Hepatitis B virus , Immunoassay , Immunoenzyme Techniques , Luminescence , Mass Screening , PlasmaABSTRACT
BACKGROUND: The Korean Red Cross (KRC) has stored blood donor samples for 10 years under -20degrees C since 2004. These samples have been used for investigating transfusion related infections and for Look-back studies. We designed an experimental scheme to verify the stability of stored blood samples. METHODS: We collected and prepared samples such as blood donor samples (HBV, HCV, HIV nucleic acid positive; n=90), the HIV infected patient samples (n=20), the WHO nucleic acid international standards serologic positive samples (HBsAg, anti-HCV, anti-HIV; n=120) and the negative samples (n=20). The samples were aliquoted in cryo tubes with volumes of 0.5~5 mL and they were stored at -20~-30degrees C and -70~-80degrees C. We used enzyme immunoassay, chemiluminescence immunoassay and quantitative PCR for the base line and the follow up studies. The linear mixed statistical model using SAS 9.1 for windows was used for statistical analysis. RESULTS: The results of the baseline test of the stored samples showed a variable range of viral load (10(1)~10(7) IU/mL or copies/mL) and optical density (S/CO 3.0~500). The results of the stored samples after 6 month (n=82) did not show any significant differences compared to the baseline data for the viral loads (P>0.05) and the qualitative serologic tests. CONCLUSION: We established an experimental scheme to verify the stability of the stored blood donor samples. From now on, the stability of the stored samples is going to be monitored by every 6 month for 10 years.