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JAIDS Journal of Acquired Immune Deficiency Syndromes:
Basic Science

Modified Dynabeads Method for Enumerating CD4+ T-Lymphocyte Count for Widespread Use in Resource-Limited Situations

Bi, Xiuqiong*†; Gatanaga, Hiroyuki*; Tanaka, Mari*; Honda, Miwako*; Ida, Setsuko*; Kimura, Satoshi*; Oka, Shinichi*

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Abstract

The Dynabeads method showed the potential for enumerating CD4+ T lymphocytes (CD4 count) in HIV-1-infected individuals. The large volume of Dynabeads required for 1 sample and complex procedure made the method expensive and not easy for use, however. To decrease the cost and simplify the procedure, we reduced the volume of the Dynabeads, added wash times, and skipped over the staining step so as to count the CD4 cells directly under an optical microscope. The CD4 count of 246 blood samples using our modified Dynabeads method (DynabeadsCD4) showed a significant correlation with that obtained by flow cytometry (FlowcytoCD4) (r = 0.91 [P < 0.0001]; slope = 1.03, intercept = −16). The sensitivity and specificity for a CD4 count less than 200 cells/μL were 79% and 94%, and for a CD4 count less than 350 cells/μL, the sensitivity and specificity were 95% and 88%, respectively. The positive and negative predictive values for a CD4 count less than 350 cells/μL were 97% and 83%, respectively. The systematic error was 8 cells/μL (95% confidence interval [CI]: 0.4-16). The cost of Dynabeads for 1 sample was less than $1.00; thus, the estimated cost per DynabeadsCD4 test is less than $3.00, including the cost of other disposable materials. Our modified method is simple, economic, and accurate enough to monitor antiretroviral therapy in resource-limited situations.

© 2005 Lippincott Williams & Wilkins, Inc.

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