Detection of anti- Pneumocystis jirovecii antibodies in human serum using a recombinant synthetic multi-epitope kexin-ba
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BRIEF REPORT
Detection of anti-Pneumocystis jirovecii antibodies in human serum using a recombinant synthetic multi-epitope kexin-based antigen Ana Luísa Tomás 1
&
Fernando Cardoso 1
&
Bruno de Sousa 2
&
Olga Matos 1
Received: 13 April 2020 / Accepted: 26 May 2020 # The Author(s) 2020
Abstract Interest in the detection of specific anti-Pneumocystis jirovecii antibodies has emerged as less-invasive alternative diagnostic approaches. Here is presented the performance of an ELISA based on a recombinant synthetic multi-epitope kexin 1 (Kex1) antigen of P. jirovecii, previously developed. Results showed that IgM anti-Kex1 levels were found significantly increased in patients with Pneumocystis pneumonia (PcP) compared with non-PcP cases (p < 0.001), allowing a diagnostic performance of PcP with a 70.8% sensitivity and a 75.0% specificity. These results suggest that this Kex1-based ELISA is a promising tool toward the serodiagnosis of PcP when the standard methods are difficult to perform. Keywords Pneumocystis . Pneumocystosis . Kexin protease . Synthetic recombinant antigen . Serological diagnosis . ELISA
Introduction Pneumocystis jirovecii pneumonia (PcP) is still the most commonly diagnosed acquired immune deficiency syndrome defining disease in Europe [1]. Likewise, the rising number of other immunocompromised patients susceptible to P. jirovecii infection [2] warrants the need for improved disease management strategies. Nowadays, PcP diagnosis still relies on microscopic visualization of the organisms or their DNA detection in specimens obtained by invasive and expensive techniques, difficult to perform in respiratory failure patients, in children, and in resource-limited settings [3, 4]. Therefore, an alternative diagnostic approach is required. The interest in serum antibodies as alternative tools for PcP diagnosis has increased since the demonstration of the humoral immunity important role in disease resolution [5–8]. Pneumocystis major surface glycoproteins
* Olga Matos [email protected] 1
Medical Parasitology Unit, Group of Opportunistic Protozoa/HIV and Other Protozoa, Global Health and Tropical Medicine, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal
2
CINEICC, Faculdade de Psicologia e de Ciências da Educação, Universidade de Coimbra, Coimbra, Portugal
(Msg), as its most abundant cell surface proteins, were the obvious candidates to start studying serological responses against PcP [9–14]. However, Msg’ variability [15] may compromise the accuracy of serological tests when recombinant antigens of this protein are used. This limitation may be the reason for the low sensitivity (68.0%) and specificity (61.8%) of a previously developed Msg-based ELISA [14]. Therefore, new antigenic candidates have been explored. Reports of high human antibodies’ titers to a recombinant subunit of Pneumocystis kexin-like serine protease (Kex1) correlated with a reduced incidence of PcP [13], protection against acquisition of Pneumocystis infection by vaccination with recomb
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