Volume 20, Number 1—January 2014
Detection of Infectivity in Blood of Persons with Variant and Sporadic Creutzfeldt-Jakob Disease
|Mouse model||Donor||Specimen||Inoculated mice||Positive mice||Incubation period, d||ID/mL (95%CI)‡|
|tgBov||vCJD||Leukocyte||24||3||476, 567, 576||2.23 (0–4.87)|
|tgHu||sCJD case 1||Plasma||14§||1||338||3.70 (0–11.65)|
|Brain||6||6||216 ± 2||NA|
|sCJD case 2||Plasma||24||0||>700||0 (0- 6.24)|
|brain||6||6||217 ± 5||NA|
|sCJD case 3||Plasma||24||1||233||2.12 (0–6.52)|
|Brain||6||6||205 ± 5||NA|
|sCJD case 4||Plasma||24||0||>700||0 (0–6.24)|
|Brain||6||6||207 ± 3||NA|
*vCJD, variant Creutzfeld-Jakob disease; sCJD, sporadic Creutzfeld-Jakob disease; dpi, days postinfection; ID, infectious dose; tgBov, bovine prion protein; tgHu, human prion protein;; PBS, phosphate-buffered saline.
†The leukocyte(s) from a single vCJD case corresponding to a starting volume of 3 mL of blood were suspended in 1 mL of 5% glucose solution. The leukocyte suspension and the crude erythrocytes were homogenized by using a high speed cell disrupter. The leukocyte and erythrocyte homogenates (vCJD case) and crude plasma (vCJD and sCJD cases) were intracerebrally injected into mice (20 µL per mouse). For the 4 sCJD MM1 cases, brain homogenate (10%, temporal cortex) were also inoculated in tgHu. Mice were euthanized when they showed clinical signs of infection or after 650 or 750 dpi. Mice were considered infected when abnormal protease-resistant prion protein; deposition was detected in brain tissue by using Western blot analysis with Sha31 monoclonal antibody: epitope amino acids 145–152 (YEDRYYRE) of the sheep PrP sequence. For samples showing 100% attack rate, incubation periods are reported as mean (± SD). For other samples, individual incubation period of CJD-positive mice are presented; their infectious titers were estimated by using limiting dilution titration method (application of Poisson model) described by Brown et al (13).
‡Leukocyte titer is expressed as ID/mL of the starting whole blood. Plasma and erythrocyte titers are expressed as ID/mL of inoculum.
§24 mice were inoculated; 10 died because of the acute toxicity of the sample.
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