rodriguez_plos05

rodriguez_plos05 - Harvard-MIT Division of Health Sciences...

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A Microchip CD4 Counting Method for HIV Monitoring in Resource-Poor Settings 1,2,3* 4 4 3 4 William R. Rodriguez , Nicolaos Christodoulides , Pierre N. Floriano , Susan Graham , Sanghamitra Mohanty , 1 1 5 5 5 4 4 Meredith Dixon , Mina Hsiang , Trevor Peter , Shabnam Zavahir , Ibou Thior , Dwight Romanovicz , Bruce Bernard , 4 1,2 4* Adrian P. Goodey , Bruce D. Walker , John T. McDevitt 1 Partners AIDS Research Center, Massachusetts General Hospital, Charlestown, Massachusetts, United States of America, 2 Division of AIDS, Harvard Medical School, Boston, Massachusetts, United States of America, 3 Brigham and Women’s Hospital, Boston, Massachusetts, United States of America, 4 Department of Chemistry and Biochemistry, University of Texas, Austin, Texas, United States of America, 5 Botswana–Harvard AIDS Institute Partnership, Princess Marina Hospital, Gaborone, Botswana Competing Interests: WRR, NC, PNF, BDW, and JTM have applied for a patent for the application de- scribed here. Author Contributions: WRR, NC, PF, SG, BDW, and JTM designed the study. WRR, NC, PNF, SG, MD, SM, ST, IB, TP, MH, DR, BB, APG, BDW, and JTM collected and analyzed the data. WRR, BDW, NC, PNF, and JTM prepared the manuscript. Academic Editor: Zvi Bentwich, Rosetta Genomics, Israel Citation: Rodriguez WR, Christo- doulides N, Floriano PN, Graham S, Mohanty S, et al. (2005) A microchip CD4 counting method for HIV mon- itoring in resource-poor settings. PLoS Med 2(7): e182. Received: January 31, 2005 Accepted: April 26, 2005 Published: July 19, 2005 DOI: 10.1371/journal.pmed.0020182 Copyright: Ó 2005 Rodriguez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abbreviations: CCD, charge- coupled device; ETC, electronic taste chip; PBS, phosphate buffered saline *To whom correspondence should be addressed. E-mail: [email protected] partners.org (WRR), [email protected] utexas.edu (JTM) A B S T R A C T Background More than 35 million people in developing countries are living with HIV infection. An enormous global effort is now underway to bring antiretroviral treatment to at least 3 million of those infected. While drug prices have dropped considerably, the cost and technical complexity of laboratory tests essential for the management of HIV disease, such as CD4 cell counts, remain prohibitive. New, simple, and affordable methods for measuring CD4 cells that can be implemented in resource-scarce settings are urgently needed. Methods and Findings Here we describe the development of a prototype for a simple, rapid, and affordable method for counting CD4 lymphocytes. Microliter volumes of blood without further sample preparation are stained with fluorescent antibodies, captured on a membrane within a miniaturized flow cell and imaged through microscope optics with the type of charge-coupled device developed for digital camera technology. An associated computer algorithm converts the raw digital image into absolute CD4 counts and CD4 percentages in real time. The accuracy of this
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