Effectiveness of Malaria Antibody Test for Screening Blood Donors / 대한수혈학회지

The Republic of Korea has been using malaria antibody assays to screen blood donors and reduce the risk of transfusion-transmitted malaria (TTM). This study examined the effectiveness of the current malaria antibody test for screening blood donors and calculated the positive predictive value (PPV) with the real-time polymerase chain reaction (RT-PCR) as the reference. The reactive rate and PPV of the malaria antibody screening assay during particular period from 2020 to 2021 were 0.82% (248/30,309) and 0.40% (1/248), respectively. The low PPV of current malaria antibody screening in blood donors suggests that the effectiveness of this test is limited in terms of balancing blood safety and supply in low-prevalence situations.

Freezing and Washing of Red Blood Cells Using Haemonetics ACP 215 / 대한수혈학회지

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.

Requirement of Establishment of Frozen Blood Storage System for Management of Rare Blood Supply and Strategic National Stockpile / 대한수혈학회지

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.

Long-Term Stability of HBV, HCV, and HIV-1 National Reference Standards for in vitro Diagnostic Medical Devices Intended to Be Used for the Nucleic Acid Amplification Test / 대한수혈학회지

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.

Evaluation of Quality of Washed Platelets Stored in Platelet Additive Solutions / 대한수혈학회지

BACKGROUND: Because of a lack of substances for platelet (PLT) metabolism and preservation, normal saline (NS) washed PLTs can only be stored for short lengths of time. However, the use of platelet additive solutions (PAS) could help solve this problem. In this study, the in vitro quality of NS washed platelets (wPLTs) stored in two types of PAS were compared with those of wPLTs stored in NS. METHODS: Five units of NS washed apheresis platelets were pooled aseptically and separated into five aliquots for storage in NS only as well as T-PAS+ (Terumo BCT, Lakewood, CO, USA) and CompoSol PS (Fenwal, Lake Zurich, IL, USA) with or without 15 mM glucose. The parameters of wPLTs quality were assessed up to 48 hrs after washing and the whole experiment was repeated 10 times independently. RESULTS: wPLTs in two kinds of PAS had better quality than wPLTs in NS, and wPLTs in T-PAS+ showed better quality than those in CompoSol PS. PAS-stored wPLTs with added glucose maintained stable CD62P and Annexin V expression during storage, but exhibited increased lactate accumulation. Evaluation of in vitro quality revealed that all wPLTs had a rating of 4 immediately after washing. However, only T-PAS+-stored wPLTs with glucose maintained a rating of 4 up to 48 hrs of post-washing. CONCLUSION: Using PAS storage for wPLTs may be beneficial compared to NS. The results presented herein suggest that T-PAS+ containing glucose has the potential to extend storage time by up to 48-hours.

Current Status of Irradiated Blood Components and Blood Irradiators in Korean Medical Institutes / 대한수혈학회지

BACKGROUND: Gamma or X-ray irradiation of blood components is used to prevent transfusion associated graft-versus-host disease (TA-GVHD). In this study, we assessed the current status of irradiated blood components and blood irradiators in Korean medical institutes. METHODS: We surveyed 306 medical institutes in Korea by a questionnaire, between August 2015 and October 2015. Institutions were asked to answer 9~16 questions, including whether they had facilities for irradiation of blood, type of irradiators used, dose of irradiation used, and if they did not have irradiation facilities, they were asked whether their blood components were irradiated. RESULTS: One-hundred and ninety-seven (64.4%) out of 306 questionnaires were returned and analyzed: 96 institutions provide irradiated blood, and 101 institutions do not use irradiated blood components. Forty-eight institutions have on site facilities with gamma blood irradiator for the irradiation of blood components and uses a dose of 20 to 50 Gy. Of the 48 institutions without facilities that use irradiated blood components, 38 (79.2%) have their blood components by referral to Korean Red Cross Blood Centers and 9 (18.8%) refer to other medical institutes for their irradiation needs. The survey showed that there is lot of regional variation in the supply and demand of irradiated blood components in Korea. CONCLUSION: Our survey does suggest that the establishment of the supply system for irradiated blood component by nation-wide blood establishments may provide national nuclear safety and stability of irradiated blood supply. It may also alleviate some regional disparity for the transfusion service of irradiated blood in Korea.

Evaluation of Two Noninvasive Hemoglobin Testing Devices / 대한수혈학회지

BACKGROUND: Determination of the hemoglobin (Hb) levels of prospective blood donors has been performed on capillary blood obtained by finger prick using a gravimetric CuSO4 method. Noninvasive Hb testing devices based on pulse oximetry technology have recently been developed. This study was conducted to evaluate the performance of two noninvasive Hb testing devices, NBM 200 and Pronto-7 as a predonation Hb screening test. METHODS: Hb levels of 993 blood donors (727 males, 266 females) were measured using five methods: two noninvasive methods, CuSO4 method, HemoCue, and hematology analyzer (Sysmex KX-21N). The hematology analyzer was considered as the reference method. RESULTS: Compared with Hb levels of the hematology analyzer, the bias was 0.7 g/dL for NBM 200, 0.1 g/dL for Prtonto-7, and 0.4 g/dL for HemoCue. The intraclass correlation coefficients of Hb measurements compared to the hematology analyzer were 0.57 (95% CI: 0.25~0.73) for NBM 200, 0.73 (95% CI: 0.69~0.75) for Pronto-7, and 0.87 (95% CI: 0.69~0.93) for HemoCue. The ability to detect Hb or =12.5 g/dL was 16.4% and 99.2% for NBM 200, 55.8% and 95.9% for Pronto-7, 60.0% and 98.6% for HemoCue and 81.8% and 95.2% for the CuSO4 method, respectively. CONCLUSION: Unsatisfactory results were obtained using the noninvasive Hb testing devices for a predonation Hb screening test, although they have the apparent advantage of reducing pain and stress in donors thereby increasing donor satisfaction. However, for application in the blood donation setting, performance of these devices should be improved.

Assessment of Serum Ferritin Levels in Plateletpheresis Donors / 대한수혈학회지

BACKGROUND: While plateletpheresis donation results in less red blood cell loss and therefore less depletion of storage iron, repeated plateletpheresis can also lead to iron depletion. To determine the safety of regular plateletpheresis donations, this study estimated donor's iron status according to age, gender, number of donations, and donation interval. METHODS: The study population included 5,109 plateletpheresis donors (4,824 males, 285 females), who passed the hemoglobin (Hb) criteria for plateletpheresis donation of 12.0 g/dL or more in an inclusion period (September 2013~November 2013). During donor screening, serum ferritin levels were measured for assessment of iron status of plateletpheresis donors. RESULTS: Mean age of donors was 30.4 years (range: 17~59). Donors with a history of donation of more than 3 years accounted for 89.3% and 74.0% in males and females, respectively. Mean donation interval and annual donation number in male (female) donors was 11.9 (7.2) weeks and 4.2 (8.7) times, respectively. Approximately 37.8% of male donors and 64.2% of female donors had a serum ferritin level of less than 15 ng/mL. Serum ferritin levels showed correlation with donation interval, as the percentage of donors with a low ferritin level decreased with increase in donation interval (rho: 0.191~0.438, P<0.001). Serum ferritin levels also showed correlation with annual plateletpheresis number (rho: -0.261~-0.411, P<0.001). CONCLUSION: Depleted iron store was observed in nearly 40% of donors who had acceptable Hb levels for plateletpheresis donation. Hb pre-donation screening is not sufficient to reduce the risk of iron deficiency in regular plateletpheresis donors.

Evaluation of Point-of-Care Testing Devices for Pre-donation Alanine Aminotransferase Test / 대한수혈학회지

BACKGROUND: In Korea, since 1990, in an effort to reduce the transmission of non-A, non-B hepatitis, all blood donations with alanine aminotransferase (ALT) levels above 65 IU/L are discarded. In 2012, 64.8% of the disposed blood units at the Korean Red Cross blood centers were due to high ALT levels. Pre-donation ALT testing might prevent unnecessary blood donation and save related expenses. We evaluated performance of point-of-care test (POCT) devices for pre-donation ALT screening. METHODS: ALT levels by four ALT POCT devices (Mission C100, Acon; Reflotron Plus, Roche; Labgeo PT10, Samsung; and FDC NX500, Fujifilm) were compared with venous blood results using laboratory chemistry analyzers (AU series, Beckman Coulter Inc.). Intraclass correlation coefficients (ICCs), sensitivity (ability to detect ALT > or =65 IU/L), and specificity (ability to detect <65 IU/L) for each method were calculated. RESULTS: Compared with the laboratory analyzers, the ICCs of ALT measurements by Mission C-100, Reflotron Plus, Labgeo PT10, and FDC NX500 were 0.96 (95% confidence interval (CI): 0.95~0.97), 0.99 (95% CI: 0.99~0.99), 0.98 (95% CI: 0.98~0.98), and 0.94 (95% CI: 0.91~0.96), respectively. The sensitivity was 80.95% for Mission C-100, 83.33% for Reflotron Plus, 78.57% for Labgeo PT10, and 97.62% for FDC NX500. The specificity was 99.13% for Mission C-100, 100.00% for Reflotron Plus, 99.78% for Labgeo PT10, and 98.26% for FDC NX500. CONCLUSION: The ALT POCT devices showed almost perfect agreement with the laboratory analyzers and could be useful for pre-donation ALT screening. However, before implementing ALT POCT devices, cost-effectiveness analyses should be performed.

Development and Preliminary Evaluation of a Leukocyte Removal Aptamer Filter / 대한수혈학회지

BACKGROUND: Leukocyte reduction filters are widely used to prevent transfusion reactions caused by leukocytes in blood components. Commercial filters are not sufficient for removal of leukocytes for prevention of transfusion associated graft-versus-host disease; therefore, irradiation of blood components was performed using expensive equipment. Techniques using an aptamer substituted for antibody have been developed and are available in clinical areas. The purpose of this study is to develop the aptamer filter system and to evaluate its efficiency and the possibility of its clinical application. METHODS: Aptamers targeted to CD45 were selected by the Postech Aptamer Initiative. The aptamer filter in which aptamers attached to beads were bound to leukocytes and removed by magnetic field was developed. Filtration of 14 units of leukoreduction-red blood provided by Korean Red Cross Blood Services was performed using aptamer filters. Leukocyte removal rate and red cell recovery rate were evaluated and bacterial culture was performed. RESULTS: After filtration using the aptamer filters, 45.6% of leukocytes were additionally removed and the red cell recovery rate was 92.8%. No growth in the bacterial culture was observed. CONCLUSION: In order to apply the cell depletion technique utilizing an aptamer to blood filter system, we developed and evaluated the aptamer filter system. Through improvement of the binding efficiency of the aptamer and the filtering process, and application of the various aptamers for other different cells, we suggest that this technique can be applied in the clinical area, such as a substitution for the irradiation process for TAGVHD prevention.

Analysis of the Coagulation Factors of Donated Plasma for Effective Utilization / 대한수혈학회지

BACKGROUND: The aim of present study was to assess the effect of different freezing time after phlebotomy on the activity of coagulation factors in frozen plasma and to evaluate which source plasma for clotting factor fractionation is appropriate for use. METHODS: Blood plasma units rejected due to a high level of ALT were divided into four groups depending on freezing time after phlebotomy, and each unit of the four groups was assayed for six different clotting factors and blood type. SAS 9.2 was used for statistical analysis of data. RESULTS: A decrease was observed in the activities of FVIII of the plasmas, in the following order: PL-A>FFP>FP(8-24)approximatelyFP(24-72). Results of the assay also showed that the levels of FVIII were significantly higher in the AB type plasmas than in the O type plasmas. PL-A and FFP units met the current quality requirements of the Korean Red Cross, in which the FVIII activity should have more than 0.7 IU/mL in more than 75% of the source plasma, as 85.0% and 82.5%, respectively. On the other hand, FP24 met the Canadian (Quebec) requirements for the source plasma, in which the FVIII activity should have more than 0.52 IU/mL in more than 75% of the source plasma, as 82.6%. CONCLUSION: For use of plasma frozen within 24 hours after phlebotomy (FP24) and plasma of specific blood type, European Pharmacopeia and WHO guidelines on quality control should be adopted for production of plasma-derived coagulation factors in Korea.

Analysis for Human Immunodeficiency Virus 1 Subtype in Korean Blood Donors / 대한수혈학회지

BACKGROUND: Genetic variants of virus appear to differ depending on the country, race, infection route, and so on. To characterize the main HIV subtype in infected blood donors and inquire about the route of HIV infection, we analyzed HIV subtype for samples that showed reactive results on the anti-HIV 1/2 and HIV-1 NAT test from September 2007 to February 2010. METHODS: To identify the HIV-1 subtype of the 90 samples that showed reactive results on the anti-HIV test and HIV-1 NAT, we performed HIV 1/2 Western blot assay, HIV RNA quantitative assay, HIV-1 nested PCR, and HIV-1 RNA sequencing. RESULTS: A total of 85 samples (94.4%) were confirmed to be HIV-1 subtypes. Among them, 82 samples (96.5%) were subtype B; and subtype A, C, and G was confirmed for one case each (1.2% for each case). We could not identify the subtype of the other five samples. One of them was amplified by nested PCR, but was not confirmed of the subtype, and four samples were not amplified even by nested PCR. CONCLUSION: The main HIV-1 subtype among the HIV-infected blood donors was confirmed to be subtype B. In addition, we identified one case each of HIV-1 subtype A, C, and G, which was not detected in blood donors in the past. It appeared that the route of HIV infection in Korea had become complicated. Therefore, we concluded that continuous research for HIV subtype analysis and efficient management of blood donors is needed.

Comparison of Methods for Detecting Bacterial Contamination in Platelet Concentrates / 대한수혈학회지

BACKGROUND: Bacterial contamination of platelets represents the highest infectious risk for a transfusion. In this study, we evaluated 2 culture-based systems that have been approved by the US FDA for bacterial screening. METHODS: Platelet concentrates were inoculated with 5 bacterial species to give a final concentration of 10(0), 10(1) and 10(2) CFU/mL. Samples for culture were taken immediately after inoculation (0 hr sample) and after 24 hrs (24 hr sample). For the BacT/ALERT 3D system, a 10 mL sample was inoculated into an aerobic culture bottle and incubated for 7 days. For the Pall eBDS system, 3 mL samples were taken from the 0 hr and 24 hr samples, respectively. The samples were incubated for 24 hrs and 30 hrs. RESULTS: Both systems detected all inoculated units both in the 0 hr and 24 hr samples, except for units inoculated with K. pneumoniae. Eleven units out of 30 units inoculated with K. pneumoniae were detected by the BacT/ALERT 3D system in the 24 hr samples. The Pall eBDS system detected 8 of 30 units in the 24 hr samples. CONCLUSION: Implementation of either system will decrease the risk of transfusing bacterially contaminated platelets. However, testing for bacterial contamination will not completely prevent septic transfusion reactions; pathogen inactivation that is now available should also be considered as an alternative method to reduce the risk of bacterial contamination.

Establishment of a Korean Hepatitis B Surface Antigen Low Titer Performance Panel for Performance Validation of Hepatitis B Surface Antigen Immunoassays / 대한수혈학회지

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.