RESUMO
The tick-borne protozoan Babesia microti is responsible for more than 200 cases of transfusion-transmitted babesiosis (TTB) infection in the United States that have occurred over the last 30 years. Measures to mitigate the risk of TTB include nucleic acid testing (NAT) and B. microti antibody testing. A fully automated prototype B. microti antibody test was developed on the Architect instrument. The specificity was determined to be 99.98% in volunteer blood donors (n = 28,740) from areas considered to have low endemicity for B. microti The sensitivity of the prototype test was studied in experimentally infected macaques; a total of 128 samples were detected as positive whereas 125 were detected as positive with an indirect fluorescent antibody (IFA) test; additionally, 83 (89.2%) of the PCR-positive samples were detected in contrast to 81 (87.1%) using an IFA test. All PCR-positive samples that tested negative in the prototype antibody test were preseroconversion period samples. Following seroconversion, periods of intermittent parasitemia occurred; 17 PCR-negative samples drawn in between PCR-positive bleed dates tested positive both by the prototype test (robust reactivity) and IFA test (marginal reactivity) prior to the administration of therapeutic drugs, indicating that the PCR test failed to detect samples from persistently infected macaques. The prototype assay detected 56 of 58 (96.6%) human subjects diagnosed with clinical babesiosis by both PCR and IFA testing. Overall, the prototype anti-Babesia assay provides a highly sensitive and specific test for the diagnosis of B. microti infection. While PCR is preferred for detection of window-period parasitemia, antibody tests detect infected subjects during periods of low-level parasitemia.
Assuntos
Anticorpos Antiprotozoários/sangue , Babesia microti/isolamento & purificação , Babesiose/diagnóstico , Imunoensaio/normas , Parasitemia/diagnóstico , Animais , Anticorpos Antiprotozoários/imunologia , Babesia microti/genética , Babesia microti/imunologia , Modelos Animais de Doenças , Técnica Indireta de Fluorescência para Anticorpo/normas , Humanos , Imunoensaio/instrumentação , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Macaca , Programas de Rastreamento , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade , Soroconversão , Reação Transfusional/prevenção & controleRESUMO
N(10)-Trifluoroacetylpteroic acid was conjugated to chemiluminescent N-sulfonylacridinium-9-carboxamide labels at the N(10) or 9-position carboxamide. Upon binding to folate binding protein the light output of the N(10) derivative (9) was quenched up to 62% upon triggering with basic peroxide, while the 9-position carboxamide conjugate (7) was quenched only 12%. The utility of this effect was demonstrated in a model homogeneous chemiluminescent assay for folic acid.
