RESUMEN
Traditional cellular and live-virus methods for detection of SARS-CoV-2 neutralizing antibodies (nAbs) are labor- and time-intensive, and thus not suited for routine use in the clinical lab to predict vaccine efficacy and natural immune protection. Here, we report the development and validation of a rapid, high throughput method for measuring SARS-CoV-2 nAbs against native-like trimeric spike proteins. This assay uses a blockade of human angiotensin converting enzyme 2 (hACE-2) binding (BoAb) approach in an automated digital immunoassay on the Quanterix HD-X platform. BoAb assays using Wuhan-WT (vaccine strain), delta (B.1.167.2), omicron BA1 and BA2 variant viral strains showed strong correlation with cell-based pseudovirus neutralization activity (PNA) and live-virus neutralization activity. Importantly, we were able to detect similar patterns of delta and omicron variant resistance to neutralization in samples with paired vaccine strain and delta variant BoAb measurements. Finally, we screened clinical samples from patients with or without evidence of SARS-CoV-2 exposure by a single-dilution screening version of our assays, finding significant nAb activity only in exposed individuals. Importantly, this completely automated assay can be performed in 4 h to measure neutralizing antibody titers for 16 samples over 8 serial dilutions or, 128 samples at a single dilution with replicates. In principle, these assays offer a rapid, robust, and scalable alternative to time-, skill-, and cost-intensive standard methods for measuring SARS-CoV-2 nAb levels.
RESUMEN
Traditional cellular and live-virus methods for detection of SARS-CoV-2 neutralizing antibodies (nAbs) are labor- and time-intensive, and thus not suited for routine use in the clinical lab to predict vaccine efficacy and natural immune protection. Here, we report the development and validation of a rapid, high throughput method for measuring SARS-CoV-2 nAbs against native-like trimeric spike proteins. This assay uses a blockade of hACE-2 binding (BoAb) approach in an automated digital immunoassay on the Quanterix HD-X platform. BoAb assays using vaccine and delta variant viral strains showed strong correlation with cell-based pseudovirus and live-virus neutralization activity. Importantly, we were able to detect similar patterns of delta variant resistance to neutralization in samples with paired vaccine and delta variant BoAb measurements. Finally, we screened clinical samples from patients with or without evidence of SARS-CoV-2 exposure by a single-dilution screening version of our assays, finding significant nAb activity only in exposed individuals. In principle, these assays offer a rapid, robust, and scalable alternative to time-, skill-, and cost-intensive standard methods for measuring SARS-CoV-2 nAb levels.
RESUMEN
Traditional cellular and live-virus methods for detection of SARS-CoV-2 neutralizing antibodies (nAbs) are labor- and time-intensive, and thus not suited for routine use in the clinical lab to predict vaccine efficacy and natural immune protection. Here, we report the development and validation of a rapid, high throughput method for measuring SARS-CoV-2 nAbs against native-like trimeric spike proteins. This assay uses a blockade of hACE-2 binding (BoAb) approach in an automated digital immunoassay on the Quanterix HD-X platform. BoAb assays using vaccine and delta variant viral strains showed strong correlation with cell-based pseudovirus and live-virus neutralization activity. Importantly, we were able to detect similar patterns of delta variant resistance to neutralization in samples with paired vaccine and delta variant BoAb measurements. Finally, we screened clinical samples from patients with or without evidence of SARS-CoV-2 exposure by a single-dilution screening version of our assays, finding significant nAb activity only in exposed individuals. In principle, these assays offer a rapid, robust, and scalable alternative to time-, skill-, and cost-intensive standard methods for measuring SARS-CoV-2 nAb levels.
RESUMEN
Attempts were made to infect 4 species of New World monkeys (Saimiri boliviensis, Aotus nancymai, A. vociferans, A. azarae boliviensis) with Plasmodium gonderi, a malaria parasite of African monkeys. Sporozoites were obtained from Anopheles dirus or A. stephensi mosquitoes that fed on an infected rhesus monkey (Macaca mulatta). Inoculation of sporozoites was by injection of dissected sporozoites by either the intravenous or intrahepatic routes, or by mosquito bite. Liver biopsies done 7 or 8 days after sporozoite inoculation showed that hepatocytes of all 4 species of these New World monkeys supported exoerythrocytic stages of P. gonderi, but daily blood film examination during a 60-day observation period failed to detect blood stages of the parasite.
Asunto(s)
Cebidae , Malaria/veterinaria , Enfermedades de los Monos/parasitología , Plasmodium/crecimiento & desarrollo , Animales , Biopsia/veterinaria , Modelos Animales de Enfermedad , Eritrocitos/parasitología , Eritrocitos/patología , Hígado/parasitología , Malaria/sangre , Malaria/parasitología , Malaria/transmisión , Enfermedades de los Monos/sangre , Enfermedades de los Monos/transmisiónRESUMEN
BACKGROUND: Although malaria is frequent in travelers, it is often misdiagnosed on initial presentation, especially in children. The objective of this study is to describe epidemiology, clinical and laboratory presentation, and treatment of children with malaria in the United States. METHODS: We performed a retrospective review of 50 confirmed cases of malaria from two pediatric metropolitan hospitals in Atlanta, GA, from 2000 to 2008. RESULTS: Malarial smears were performed in 385 unique patients; 50 (12.6%) were positive. American children who had visited family and friends in malaria-endemic countries comprised 62% of our cases. Most cases visited Nigeria or Cameroon; all but three traveled to Africa. Three patients presented 8 to 12 months following travel. Plasmodium falciparum was diagnosed most frequently (72%). Most patients had low-level parasitemia (<1%). Gametocytes were rarely identified. Treatment was primarily with quinine and either doxycycline or clindamycin, and transfusion was rare. All patients responded rapidly to treatment. Although seven (14%) had hyperparasitemia (> 5%), no fatalities or long-term sequelae were seen. CONCLUSIONS: Malarial diagnosis can be difficult in children because parasitemia is usually below 1%. A high index of suspicion is required in patients who have traveled to Africa.
Asunto(s)
Malaria/diagnóstico , Malaria/epidemiología , Parasitemia/diagnóstico , Parasitemia/epidemiología , Plasmodium falciparum/aislamiento & purificación , África , Portador Sano/parasitología , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Georgia , Hospitales Pediátricos/organización & administración , Humanos , Malaria Falciparum/diagnóstico , Malaria Falciparum/epidemiología , Estudios Retrospectivos , Factores de Riesgo , ViajeRESUMEN
We describe the 11th case of bioterrorism-related inhalational anthrax reported in the United States. The presenting clinical features of this 94-year-old woman were subtle and nondistinctive. The diagnosis was recognized because blood cultures were obtained prior to administration of antibiotics, emphasizing the importance of this diagnostic test in evaluating ill patients who have been exposed to Bacillus anthracis. The patient's clinical course was characterized by progression of respiratory insufficiency, pleural effusions and pulmonary edema, and, ultimately, death. Although her B anthracis bacteremia was rapidly sterilized after initiation of antibiotic therapy, viable B anthracis was present in postmortem mediastinal lymph node specimens. The source of exposure to B anthracis in this patient is not known. Exposure to mail that was cross-contaminated as it passed through postal facilities contaminated with B anthracis spores is one hypothesis under investigation.