Browsing by Author "Wilhelm, Jan"

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    Comparison of VSV Pseudovirus and Focus Reduction Neutralization Assays for Measurement of Anti-Andes orthohantavirusNeutralizing Antibodies in Patient Samples
    (2020) Vial, Cecilia; Whitaker, Annalis; Wilhelm, Jan; Ovalle, Jimena; Perez, Ruth; Valdivieso, Francisca; Ferres, Marcela; Martinez-Valdebenito, Constanza; Eisenhauer, Philip; Mertz, Gregory J.; Hooper, Jay W.; Botten, Jason W.; Vial, Pablo A.
    Andes orthohantavirus(ANDV) is the etiologic agent of hantavirus cardiopulmonary syndrome (HCPS), which has a case fatality rate around 35%, with no effective treatment or vaccine available. ANDV neutralizing antibody (NAb) measurements are important for the evaluation of the immune response following infection, vaccination, or passive administration of investigational monoclonal or polyclonal antibodies. The standard assay for NAb measurement is a focus reduction neutralization test (FRNT) featuring live ANDV and must be completed under biosafety level (BSL)-3 conditions. In this study, we compared neutralization assays featuring infectious ANDV or vesicular stomatitis virus (VSV) pseudovirions decorated with ANDV glycoproteins for their ability to measure anti-ANDV NAbs from patient samples. Our studies demonstrate that VSV pseudovirions effectively measure NAb from clinical samples and have greater sensitivity compared to FRNT with live ANDV. Importantly, the pseudovirus assay requires less labor and sample materials and can be conducted at BSL-2.
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    Evaluación de la sensibilidad diagnóstica de tres técnicas de laboratorio para la infección por influenza A: inmunocromatografía, IFD e IFD con citocentrifugado versus RPC-TR
    (SOC CHILENA INFECTOLOGIA, 2014) Wilhelm, Jan; Yubero, Joao; Fuentes, Constanza; Rios, Paulina; Leyton, Oscar; Reyes Zaldívar, Felipe Tomas
    Introduction: The specific diagnosis of influenza A infection makes. it possible to control its spread, decreases the unnecessary use of antibiotics, clinical procedures and laboratory test, and allows early recognition of outbreaks. Different technologies are currently available in Chile for this purpose. Objective: The study presented here compares the sensitivity for influenza A virus detection of immunocromatography (RIDT), direct fluorescent antibodies-DFA and DFA with cytocentrifugation against the gold standard, RT-PCR. Material and Methods: In 175 nasal swab samples influenza, RIDT and RT-PCR were performed. Another 1689 nasal swab samples were tested by DFA and RT-PCR for influenza A. Finally, 29 nasal swab samples confirmed as Influenza A positive by RT-PCR welt tested by DFA with cytocentrifugation. Results: The RIDT, DFA and DFA + cytocentrifugation sensitivity was 47,3%, 57,2% and 72,4%, respectively. Discussion and Conclusion: Their lower cost and faster turnaround time when compared to PCR make RIDT and DFA the tests of choice in diagnostic laboratories in Chile. However, their low sensitivity and NPV, especially during low season, makes more sensitive diagnostic tools necessary to confirm the results. In our study cytocentrifugation increased DFA sensitivity from 57% to 72%.