Multisite Validation of Cryptococcal Antigen Lateral Flow Assay and Quantification by Laser Thermal Contrast
David R. Boulware

, Melissa A. Rolfes, Radha Rajasingham, Maximilian von Hohenberg, Zhenpeng Qin, Kabanda Taseera, Charlotte Schutz, Richard Kwizera, Elissa K. Butler, Graeme Meintjes, Conrad Muzoora, John C. Bischof, and David B. Meya
Author affiliations: University of Minnesota, Minneapolis, Minnesota, USA (D.R. Boulware, M.A. Rolfes, R. Rajasingham, M. von Hohenberg, Z. Qin, E.K. Butler, J.C. Bischof, D.B. Meya); Makerere University, Kampala, Uganda (R. Rajasingham, R.K. Kwizera, D.B. Meya); Mbarara University of Science and Technology, Mbarara, Uganda (K. Taseera, C. Muzoora); G.F. Jooste Hospital, Cape Town, South Africa (C. Schutz, G. Meintjes); University of Cape Town, Cape Town (C. Schutz, G. Meintjes)
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Figure 3
Figure 3. . A) Prediction of cryptococcal antigen titer based on laser thermal contrast measurement and concept of lateral flow immunochromatographic assay (LFA) thermal contrast measurement in which a laser irradiates the test line in the LFA (19). The test line is formed by gold–monoclonal antibody–antigen sandwich complex with a monoclonal antibody affixed at the test line. When irradiated by a green laser (532 nm), any gold present absorbs light from the laser and generates heat in direct proportion to the amount of gold (and thereby antigen) present at the test line. This temperature change can be measured by using an infrared camera. B) Association of measured semiquantitative LFA cryptococcal antigen (CRAG) titer starting at a 1:250 dilution by the predicted CRAG titer based on thermal contrast measurement. Measurements on the negative portion of the x-axis are beyond the visual range when specimens were diluted 1:250, yet still detectable by thermal contrast. The Pearson correlation coefficient was r = 0.91 (p<0.001, R2 = 0.84) among 115 positive specimens quantified. A total of 58 LFA CRAG–negative specimens established background levels of heat radiation.
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