A sensitive and quantitative assay for normal PrP in plasma. Export

@article{citeulike:2894781, abstract = {Transmissible spongiform encephalopathies can be transmitted by blood transfusion. The risk of spreading the disease among the human population could be mitigated with the implementation of a blood screening assay. We developed a two-antibody assay for PrP detection in plasma using the ORIGEN technology with a protocol modification to improve the limit of detection and to increase the sample volume assayed. In the standard 200 microL format, the assay had a detection limit of 7-10 pg of recombinant PrP and 3 pg in 1 mL final volume implementation. PrP concentration measured in normal and scrapie-infected hamster brains was 7.5+/-0.9 and 57.3+/-9.6 microg/g, respectively. After a concentration step with an immuno-affinity resin, plasma PrP(c) was detected by Western blot and its concentration was measured at 3.5+/-0.8 ng/mL. From these data and assuming that blood has the same specific infectivity as brain, we estimated the concentration of abnormal PrP in hamster-infected plasma to be 32 f g/mL. The assay also detected abnormal brain PrP spiked into plasma although the limit of detection was affected. This is a novel and sensitive assay for the detection of PrP in plasma that could be developed into a platform for a plasma-based TSE test.}, address = {Veterans Affairs Medical Center and University of Maryland, Baltimore, MD 21201, United States. lgreg002@umaryland.edu}, author = {Gregori, L. and Gray, B. N. and Rose, E. and Spinner, D. S. and Kascsak, R. J. and Rohwer, R. G.}, citeulike-article-id = {2894781}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.jviromet.2008.01.028}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/18339433}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=18339433}, doi = {10.1016/j.jviromet.2008.01.028}, issn = {0166-0934}, journal = {Journal of virological methods}, keywords = {diagnosis}, month = {May}, number = {2}, pages = {251--259}, posted-at = {2008-06-14 16:18:37}, priority = {5}, title = {A sensitive and quantitative assay for normal PrP in plasma.}, url = {http://dx.doi.org/10.1016/j.jviromet.2008.01.028}, volume = {149}, year = {2008} }

TY - JOUR ID - citeulike:2894781 L3 - citeulike-article-id:2894781 N2 - Transmissible spongiform encephalopathies can be transmitted by blood transfusion. The risk of spreading the disease among the human population could be mitigated with the implementation of a blood screening assay. We developed a two-antibody assay for PrP detection in plasma using the ORIGEN technology with a protocol modification to improve the limit of detection and to increase the sample volume assayed. In the standard 200 microL format, the assay had a detection limit of 7-10 pg of recombinant PrP and 3 pg in 1 mL final volume implementation. PrP concentration measured in normal and scrapie-infected hamster brains was 7.5+/-0.9 and 57.3+/-9.6 microg/g, respectively. After a concentration step with an immuno-affinity resin, plasma PrP(c) was detected by Western blot and its concentration was measured at 3.5+/-0.8 ng/mL. From these data and assuming that blood has the same specific infectivity as brain, we estimated the concentration of abnormal PrP in hamster-infected plasma to be 32 f g/mL. The assay also detected abnormal brain PrP spiked into plasma although the limit of detection was affected. This is a novel and sensitive assay for the detection of PrP in plasma that could be developed into a platform for a plasma-based TSE test. IS - 2 JF - Journal of virological methods SN - 0166-0934 AD - Veterans Affairs Medical Center and University of Maryland, Baltimore, MD 21201, United States. lgreg002@umaryland.edu EP - 259 TI - A sensitive and quantitative assay for normal PrP in plasma. VL - 149 SP - 251 KW - diagnosis AU - Gregori, L AU - Gray, BN AU - Rose, E AU - Spinner, DS AU - Kascsak, RJ AU - Rohwer, RG PY - 2008/05// UR - http://dx.doi.org/10.1016/j.jviromet.2008.01.028 ER -