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KMID : 0387720070180030194
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Korean Journal of Blood Transfusion
2007 Volume.18 No. 3 p.194 ~ p.201
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Measurement of the Residual Leukocyte Count by Flow Cytometry in Leukocyte-reduced Blood Components.
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Lee Won-Mok
Kim Ji-Hae
Ryoo Nam-Hee
Ha Jung-Sook
Jeon Dong-Suk
Kim Jae-Ryong
Park Soo-Jin
Kim Eun-Jin
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Abstract
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Background: Counting methods for residual leukocytes in leukocyte-reduced blood components include the manual assay using a Nageotte hemocytometer, flow cytometry (FCM) and polymerase chain reaction (PCR). The Nageotte hemocytometer is the most widely used technique in Korea, but it is time consuming and dependent upon the skill of the examiner. We examined residual leukocytes count using a hemocytometer and FCM and compared the accuracy of the use of both methods.
Methods: Twenty units of RBC concentrates (RCs) were collected in quadruple top-to-bottom collection bags of the Green Cross (Green Cross, Eumsung, Korea) and from BD (Becton Dickinson Korea, Gumi, Korea). Thirty-five units of single donor platelets (SDPs) collected by Amicus (Baxter, Deerfield, IL USA), MCS+ (Haemonetics, Braintree, MA, USA) and Trima (Gambro BCT, Lakewood, CO, USA) were used to measure residual leukocytes using a hemocytometer and FCM using the LeucoCOUNT system (BD Bioscience, San Jose, CA USA).
Results: Linearity of the LeucoCOUNT system was well maintained (R2=0.9998). Reproducibility was CV of 5.08% for 53.01/microlitermicroliter and 8.54% for 4.97/microliter. The hemocytometer measured lower leukocyte counts in all RCs and SDPs than the LeucoCOUNT system. The mean residual leukocyte counts in RCs measured by the hemocytometer were 0.075x10(6)/unit with the Green Cross collection bags and 0.067x10(6)/unit with the BD collection bags, but were 2.301x10(6)/unit and 1.943x10(6)/unit, respectively, when measured with use of the LeucoCOUNT system. The mean residual leukocyte count in the SDPs collected by Amicus, MCS+ and Trima, as measured by the hemocytometer, were 0.037x10(6)/unit, 0.046x10(6)/unit and 0.043x10(6)/unit, respectively. The mean residual leukocyte count in the SDPs collected by Amicus, MCS+ and Trima, as measured by the LeucoCOUNT system, were 0.061x10(6)/unit, 0.161x10(6)/unit and 0.045x10(6)/unit, respectively.
Conclusion: The use of the flow cytometric method is moresensitive and produces an accurate leukocyte count in leukocyte-reduced blood components than the use of themanual method.
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KEYWORD
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Flow cytometry, Nageotte hemocytometer, Residual leukocyte, Blood components
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