Global gene expression profile and functional analysis reveal the conservation of reproduction-associated gene networks in Gossypium hirsutum.

KEY MESSAGE: Lastly, the bZIP gene family encompasses genes that have been reported to play a role in flower development, such as bZIP14 (FD). Notably, bZIP14 is essential for Flowering Locus T (FT) initiation of floral development in Arabidopsis (Abe et al. 2005). Cotton (Gossypium hirsutum L.) is the world's most extensively cultivated fiber crop. However, its reproductive development is poorly characterized at the molecular level. Thus, this study presents a detailed transcriptomic analysis of G. hirsutum at three different reproductive stages. We provide evidence that more than 64,000 genes are active in G. hirsutum during flower development, among which 94.33% have been assigned to functional terms and specific pathways. Gene set enrichment analysis (GSEA) revealed that the biological process categories of floral organ development, pollen exine formation, and stamen development were enriched among the genes expressed during the floral development of G. hirsutum. Furthermore, we identified putative Arabidopsis homologs involved in the G. hirsutum gene regulatory network (GRN) of pollen and flower development, including transcription factors such as WUSCHEL (WUS), INNER NO OUTER (INO), AGAMOUS-LIKE 66 (AGL66), SPOROCYTELESS/NOZZLE (SPL/NZZ), DYSFUNCTIONAL TAPETUM 1 (DYT1), ABORTED MICROSPORES (AMS), and ASH1-RELATED 3 (ASHR3), which are known crucial genes for plant reproductive success. The cotton MADS-box protein-protein interaction pattern resembles the previously described patterns for AGAMOUS (AG), SEEDSTICK (STK), SHATTERPROOF (SHP), and SEPALLATA3 (SEP3) homolog proteins from Arabidopsis. In addition to serving as a resource for comparative flower development studies, this work highlights the changes in gene expression profiles and molecular networks underlying stages that are valuable for cotton breeding improvement.

Viability and oxidative stress of dental pulp cells after indirect application of chemomechanical agents: An in vitro study.

AIM: This study evaluated the transdentinal cytotoxic effects of enzymatic agents (EA) for chemomechanical carious tissue removal on human dental pulp cells. METHODOLOGY: The groups were based on the performed dentine treatments (n = 8): G1: Positive Control (PC - no treatment); G2: Negative Control (NC - 35% H2 O2 for 2 min); G3: Brix 3000™ (BX) for 30 s; G4: BX for 2 min; G5: Papacarie Duo™ (PD) for 30 s; G6: PD for 2 min. The cells were evaluated for viability (VB; MTT assay) and production of reactive oxygen species (ROS; DCFH-DA assay) and nitric oxide (NO; Griess reagent). A scanning electron microscope provided morphological chemical analyses and energy-dispersive X-ray spectroscopy. The data were submitted to the one-way anova statistical test complemented by Tukey (p < .05). RESULTS: Cell viability decreased by 21.1% and 58.4% in G5 and G6, respectively. ROS production in G3 and G4 maintained basal levels but increased by 171.2% and 75.1% in G5 and G6, respectively. CONCLUSIONS: The Brix3000™ enzymatic agent did not cause indirect cytotoxic effects on pulp cells, regardless of the application time. Conversely, Papacarie Duo™ reduced viability and increased ROS production by pulp cells.

Práticas extensionistas sob a perspectiva teórica das universidades promotoras da saúde / Prácticas de extensión desde la perspectiva teórica de las universidades promotoras de salud / Extension practices from the theoretical perspective of health-promoting universities

Objetivo:analisar relações existentes entre proposta conceitual do movimento das Universidades Promotoras da Saúde e projetos de extensão de uma universidade, sob perspectiva de uma análise documental. Método:estudo de caso documental com análise qualitativa descritiva realizado com base nos documentos de registros extensionistas de uma universidade pública do estado do Rio de Janeiro. A análise deste estudo teve como aporte teórico a proposta conceitual do movimento das Universidades Promotoras da Saúde. Resultados:identificaram-se 11 projetos que atenderam aos critérios de inclusão e, foram distribuídos em cinco categorias. Conclusão:é necessário vincular a promoção da saúde ao projeto político pedagógico da Universidade, possibilitando a valorização das experiências formativas, no âmbito das histórias de vida dos sujeitos e das suas vivências comunitárias e no âmbito da política institucional. Entretanto, tais iniciativas ainda são escassas, gerando um hiato entre expectativa e realidade.

Objective:to analyze existing relationships between the conceptual proposal of the health promoting universities movement and university extension projects, from the perspective of a documental analysis. Method:documental case study with descriptive qualitative analysis based on records documents from a public university in the state of Rio de Janeiro. The analysis of this study had as theoretical contribution the conceptual proposal of the Health Promoting Universities movement. Results: 11projects thatmet the inclusion criteria were identified and distributed into five categories. Conclusion:it is necessary to link health promotion to the University's political pedagogical project, enabling the valorization of formative experiences, within the scope of the subjects' life stories, their community experiences and within the scope of institutional policy. However, such initiatives are still scarce, creating a gap between expectations and reality.

Objetivo:analizar relaciones existentes entre propuesta conceptual del movimiento Universidades Promotoras de Salud y los proyectos de extensión de una universidad, desde la perspectiva de un análisis documental. Método:estudio de caso documental con análisis cualitativo descriptivo basado en registros extensión de una universidad pública del estado de Río de Janeiro. El análisis de este estudio tuvo como soporte teórico del movimiento de Universidades Promotoras de Salud. Resultados:se identificaron once proyectos que cumplieron con los criterios de inclusión y se distribuyeron en cinco categorías. Conclusión:es necesario vincular la promoción de la salud al proyecto político pedagógico de la Universidad, posibilitando la valorización de las experiencias formativas, en el ámbito de las historias de vida de los sujetos y de sus experiencias comunitarias y en elámbito de la política institucional. Sin embargo, tales iniciativas aún son escasas, creando una brecha entre las expectativas y la realidad.

The chloroplast protein HCF164 is predicted to be associated with Coffea SH9 resistance factor against Hemileia vastatrix.

To explore the connection between chloroplast and coffee resistance factors, designated as SH1 to SH9, whole genomic DNA of 42 coffee genotypes was sequenced, and entire chloroplast genomes were de novo assembled. The chloroplast phylogenetic haplotype network clustered individuals per species instead of SH factors. However, for the first time, it allowed the molecular validation of Coffea arabica as the maternal parent of the spontaneous hybrid "Híbrido de Timor". Individual reads were also aligned on the C. arabica reference genome to relate SH factors with chloroplast metabolism, and an in-silico analysis of selected nuclear-encoded chloroplast proteins (132 proteins) was performed. The nuclear-encoded thioredoxin-like membrane protein HCF164 enabled the discrimination of individuals with and without the SH9 factor, due to specific DNA variants linked to chromosome 7c (from C. canephora-derived sub-genome). The absence of both the thioredoxin domain and redox-active disulphide center in the HCF164 protein, observed in SH9 individuals, raises the possibility of potential implications on redox regulation. For the first time, the identification of specific DNA variants of chloroplast proteins allows discriminating individuals according to the SH profile. This study introduces an unexplored strategy for identifying protein/genes associated with SH factors and candidate targets of H. vastatrix effectors, thereby creating new perspectives for coffee breeding programs.

Elevated CO2 increases biomass of Sorghum bicolor green prop roots under drought conditions via soluble sugar accumulation and photosynthetic activity.

Elevated [CO2 ] (E[CO2 ]) mitigates agricultural losses of C4 plants under drought. Although several studies have described the molecular responses of the C4 plant species Sorghum bicolor during drought exposure, few have reported the combined effects of drought and E[CO2 ] (E[CO2 ]/D) on the roots. A previous study showed that, among plant organs, green prop roots (GPRs) under E[CO2 ]/D presented the second highest increase in biomass after leaves compared with ambient [CO2 ]/D. GPRs are photosynthetically active and sensitive to drought. To understand which mechanisms are involved in the increase in biomass of GPRs, we performed transcriptome analyses of GPRs under E[CO2 ]/D. Whole-transcriptome analysis revealed several pathways altered under E[CO2 ]/D, among which photosynthesis was strongly affected. We also used previous metabolome data to support our transcriptome data. Activities associated with photosynthesis and central metabolism increased, as seen by the upregulation of photosynthesis-related genes, a rise in glucose and polyol contents, and increased contents of chlorophyll a and carotenoids. Protein-protein interaction networks revealed that proliferation, biogenesis, and homeostasis categories were enriched and contained mainly upregulated genes. The findings suggest that the previously reported increase in GPR biomass of plants grown under E[CO2 ]/D is mainly attributed to glucose and polyol accumulation, as well as photosynthesis activity and carbon provided by respiratory CO2 refixation. Our findings reveal that an intriguing and complex metabolic process occurs in GPRs under E[CO2 ]/D, showing the crucial role of these organs in plant drought /tolerance.

Aeromonas allosaccharophila Strain AE59-TE2 Is Highly Antagonistic towards Multidrug-Resistant Human Pathogens, What Does Its Genome Tell Us?

Multidrug-resistant bacteria are of critical importance and a problem for human health and food preservation; the discovery of new antimicrobial substances to control their proliferation is part of the solution. This work reports on 57 antagonistic Aeromonas strains, of which 38 strains were antagonistic towards problematic human pathogens. The genome of the most antagonistic strain was sequenced and identified as Aeromonas allosaccharophila. Its genome was fully annotated and mined for genes that might explain that activity. Strain AE59-TE was antagonistic toward clinically relevant gram-negative and gram-positive multidrug-resistant bacteria, including Klebsiella pneumoniae KPC, Escherichia coli ESBL, Salmonella typhimurium, and Staphylococcus aureus MRSA. Strain AE59-TE2 was identified by multilocus sequence analysis. Genome mining identified four genes homologous to the bacteriocin, zoocin A from Streptococcus equi and a gene 98% similar to cvpA linked to colicin V production. A. allosaccharophila strain AE59-TE2 produced antimicrobial activity against a broad range of bacteria, including important gram-negative bacteria, not typically targeted by bacteriocins. Herewere described novel zoocin genes that are promising for industrial applications in the food and health sectors. Interesting and important antagonistic activity is described combined with the first detailed genomic analysis of the species Aeromonas allosaccharophila.

Rabies virus isolated from insectivorous bats induces different inflammatory responses in experimental model.

Rabies virus (RABV) is a neurotropic virus that causes fatal neuroinflammation in mammals. The insectivorous bat RABV strains are less pathogenic for mice than strains associated with other reservoirs. We characterized the tissue inflammatory response in the CNS of RABV isolated from insectivorous bats. Eptesicus furinalis (EPBRV)-infected mice had a robust inflammatory response and a greater amount of IL-1ß, IL-6 and TNF-α, while Myotis nigricans (MNBRV)-infected mice showed a higher expression of IL-17 and greater activation of IFN-ß. New approaches to understand the inflammatory response to different mechanisms of action may provide insights for the development of novel therapies for rabies.

A comparative review of serological assays for the detection of rabies virus-specific antibodies.

Rabies is a major public health problem with a fatality rate close to 100%, caused by a virus of the Lyssavirus genus, of which rabies virus (RABV) is the prototype. Nonetheless, the complete prevention can be achieved by the induction of neutralizing antibodies by pre- or post-exposure prophylaxis. According to the world health organization (WHO) and World Organization for animal health (OIE), serum titers of rabies virus neutralizing antibodies (RVNA) that are higher or equal to 0.5 international units (IU)/ml indicate adequate immune response after vaccination against rabies. Currently, RFFIT and FAVN are the gold standard tests recommended by both WHO and OIE for detecting and quantitating RVNA in biological samples from individuals or animals previously vaccinated and/or subjects suspected of having been infected by RABV. Although the tests RFFIT and FAVN are efficient, they are time-consuming, labor-intensive manual tests and not cost-effective for routine use. Following the previously mentioned, approaches with alternative methods have been developed to detect RVNA or rabies-specific antibodies in human or animal serum, but with variable success. This work summarizes the advances in the serological assays for the detection of neutralizing antibodies or rabies antibodies and assesses the individual immune status after vaccination against rabies, as well as the mechanisms of RABV neutralization mediated by antibodies. Therefore, the main alternative methods for the determination of RABV or rabies-specific antibodies are exposed, with promising results, besides being easy to execute, of low cost, and representing a possibility of being applied, according to the proposal of each test to the network of Rabies Surveillance Laboratories.

Rabies virus isolated from insectivorous bats induces different inflammatory responses in experimental model

Rabies virus (RABV) is a neurotropic virus that causes fatal neuroinflammation in mammals. The insectivorous bat RABV strains are less pathogenic for mice than strains associated with other reservoirs. We characterized the tissue inflammatory response in the CNS of RABV isolated from insectivorous bats. Eptesicus furinalis (EPBRV)-infected mice had a robust inflammatory response and a greater amount of IL-1β, IL-6 and TNF-α, while Myotis nigricans (MNBRV)-infected mice showed a higher expression of IL-17 and greater activation of IFN-β. New approaches to understand the inflammatory response to different mechanisms of action may provide insights for the development of novel therapies for rabies.

Rabies virus isolated from insectivorous bats induces different inflammatory responses in experimental model

Rabies virus (RABV) is a neurotropic virus that causes fatal neuroinflammation in mammals. The insectivorous bat RABV strains are less pathogenic for mice than strains associated with other reservoirs. We characterized the tissue inflammatory response in the CNS of RABV isolated from insectivorous bats. Eptesicus furinalis (EPBRV)-infected mice had a robust inflammatory response and a greater amount of IL-1β, IL-6 and TNF-α, while Myotis nigricans (MNBRV)-infected mice showed a higher expression of IL-17 and greater activation of IFN-β. New approaches to understand the inflammatory response to different mechanisms of action may provide insights for the development of novel therapies for rabies.

Cytotoxicity and dentin composition alterations promoted by different chemomechanical caries removal agents: A preliminary in vitro study.

BACKGROUND: The use of chemomechanical agents for caries removal has been indicated as a non-invasive treatment option; however, their possible deleterious effects on the dental-pulp complex have been insufficiently studied. This study assessed the direct cytotoxicity of two chemomechanical caries removal agents (Brix 3000™ - BX and Papacarie Duo™ - PD) on pulp cells from deciduous teeth, as well as to assess the morphology and chemical compositions of the dentin surface after the application of these materials. MATERIAL AND METHODS: The cells were seeded (50,000 cells/cm²) in a culture medium (DMEM with 10% fetal bovine serum - FBS). After 24 hours, the BX and PD materials were added to 1:20, 1:100, and 1:1000 dilutions. Non-exposed cells were considered as the control group. The viability test (MTT), Trypan Blue assay (TB), and cell morphology (Scanning Electron Microscopy - SEM) were performed after 24 hours of agent application. For the SEM and chemical (energy-dispersive X-ray spectrometry - EDS) dentin evaluation, 0.3-mm-thick dentin discs were obtained and divided into control group (no treatment) and surfaces covered with 37% phosphoric acid, BX, or PD. Data were compared by one-way ANOVA and Tukey's test (p<0.05). RESULTS: Decreases in cell viability and numbers of viable cells were observed for both materials, at all dilutions, when compared with the control group (p<0.05). The BX and PD materials did not cause visually perceptible changes, according to SEM, on the surfaces of dentin discs. The EDS analysis did not indicate a statistically significant difference in the levels of calcium (Ca) and phosphorus (P) between the materials and the control group (p>0.05). CONCLUSIONS: Both materials showed cytotoxicity when in direct contact with the pulp cells from deciduous teeth, and the BX material presented lower cytotoxicity than the PD material. Moreover, both materials did not significantly change the dentin composition. Key words:Cell culture, cytotoxicity, dental pulp, papacarie, primary teeth.

Glycosylation is required for the neutralizing activity of human IgG1 antibodies against human rabies induced by pre-exposure prophylaxis.

Rabies lyssavirus (RABV) neutralizing IgG antibodies confer protection after rabies vaccination, although how the RABV-specific antibodies neutralize the virus is still unknown. As changes in the antibody's carbohydrate chain can interfere with its effector functions, we compared the glycosylation patterns of both neutralizing and non-neutralizing IgG1 induced by pre-exposure prophylaxis to human rabies and analyzed their influence on in vitro antibody neutralizing activities. Specific IgG1 were purified from human serum using affinity chromatography. Purity and avidity were analyzed by SDS-PAGE and indirect ELISA using NH4SCN respectively. The N-linked oligosaccharide chain of the purified IgG antibody was evaluated using a lectin-based ELISA assay with a panel of seven lectins. The activity of purified IgG1 and neutralizing IgG1 deglycosylated by PNGase F enzyme were analyzed using the rapid fluorescent focus inhibition test. The purified IgG1 showed an electrophoretic pattern compatible with human IgG. All of the antibodies recognized RABV, although neutralizing IgG1 had a higher avidity (RAI = 80%) than non-neutralizing IgG1 (RAI = 30%). The neutralizing IgG1 also showed higher binding to WFA, ECA, WGA, and ConA lectins, indicating possible different N-acetylgalactosamine, galactose, N-acetylglucosamine, and mannose contents. Non-neutralizing IgG1, on the other hand, showed strong binding at UEA-1 and SNA, which bind to fucose and sialic acid residues respectively. Different glycosylation profiles were also observed in Fab and Fc fragments from neutralizing and non-neutralizing IgG1, although the deglycosylated IgG1 lost its neutralizing activity. Our results suggest that antibody glycosylation is important for neutralizing RABV in vitro, since neutralizing IgG1 has a different glycosylation profile than non-neutralizing IgG1. Further research will be needed to better evaluate the differential glycosylation patterns between IgG1 antibodies following vaccination.

Repercussões para saúde mental de profissionais de enfermagem atuantes no enfrentamento à Covid-19: revisão integrativa / Repercussions for mental health of nursing professionals who are in the face of The Covid-19: integrative review / Repercusiones para la salud mental de los profesionales de enfermería que trabajan en el afrontamiento del Covid-19: una revisión integradora

Objetivo: identificar as repercussões na saúde mental de profissionais de enfermagem atuantes no enfrentamento da Covid-19 no primeiro ano do contexto pandêmico. Método: revisão integrativa de literatura realizada em maio de 2021 nas bases de dados eletrônicas SCOPUS, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Web of Science e Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS). Resultados: foram encontradas 1249 publicações cuja aplicação de critérios de inclusão, exclusão e leitura crítica, culminou na análise de 18 artigos provenientes de vivências da China, Turquia, Itália, Canadá, Estados Unidos, Filipinas, Brasil, Portugal e Etiópia. As principais repercussões envolvidas no adoecimento mental foram: ansiedade, depressão, estresse, síndrome de Burnout, transtornos mentais do sono e transtorno de estresse pós-traumático. Conclusão: a saúde mental dos enfermeiros encontra-se extremamente fragilizada frente ao contexto pandêmico. Essa emergência na saúde mundial, acabou por desencadear consequências físicas, emocionais e psicológicas nesses trabalhadores.

Objective: To identify the repercussions on mental health of nursing professionals who are in the face of Covid-19 in the first year of the pandemic context. Method: integrative literature review conducted in May 2021 in the electronic databases SCOPUS, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Web of Science and Latin American and Caribbean Literature on Health Sciences (LILACS). Results: 1,249 publications were found whose application of inclusion, exclusion and critical reading criteria culminated in the analysis of 18 articles from experiences from China, Turkey, Italy, Canada, United States, Philippines, Brazil, Portugal and Ethiopia. The main repercussions involved in mental illness were anxiety, depression, stress, Burnout syndrome, mental sleep disorders and posttraumatic stress disorder. Conclusion: The mental health of nurses is extremely fragile in the face of the pandemic context. This emergency in world health eventually triggered physical, emotional and psychological consequences in these workers.

Objetivo: identificar las repercusiones en la salud mental de los profesionales de enfermería que trabajan en el afrontamiento del Covid-19 en el primer año del contexto pandémico. Método: revisión integrativa de la literatura realizada en mayo de 2021 en las bases de datos electrónicas SCOPUS, Índice Acumulativo de Literatura de Enfermería y Afines en Salud (CINAHL), Web of Science y Literatura de Ciencias de la Salud de América Latina y el Caribe (LILACS). Resultados: se encontraron 1249 publicaciones cuya aplicación de criterios de inclusión, exclusión y lectura crítica culminó en el análisis de 18 artículos de experiencias en China, Turquía, Italia, Canadá, Estados Unidos, Filipinas, Brasil, Portugal y Etiopía. Las principales repercusiones de la enfermedad mental fueron: ansiedad, depresión, estrés, síndrome de Burnout, trastornos del sueño mental y trastorno por estrés postraumático. Conclusión: la salud mental de las enfermeras es extremadamente frágil frente al contexto pandémico. Esta emergencia sanitaria mundial acabó provocando consecuencias físicas, emocionales y psicológicas para estos trabajadores.

Development of biotinylated polyclonal anti-ribonucleoprotein IgG for detection of rabies virus antigen by direct rapid immunohistochemical test.

The direct rapid immunohistochemical test (dRIT) has been recommended for laboratorial diagnosis of rabies, especially in developing countries. The absence of commercial primary antibodies, however, still represents a major limitation to its wider use in testing. We describe here the development of a biotinylated polyclonal antibody against Rabies lyssavirus (RABV) ribonucleoprotein (RNP) and its use as a primary reagent in dRIT. Anti-RNP polyclonal horse IgG was purified by ionic exchange chromatography followed by immunoaffinity column chromatography, and its affinity, diagnostic sensitivity, and specificity were evaluated. CNS samples (120) of suspected rabies cases in different animal species were tested by dRIT, with the positive (n = 14) and negative (n = 106) results confirmed by direct fluorescence antibody testing (dFAT). Comparing the results of dRIT and dFAT, we found that the biotinylated anti-RNP IgG delivered 100% diagnostic specificity and sensibility for rabies diagnosis. Our findings show that the biotinylated anti-RNP polyclonal IgG can be produced with the quality required for application in dRIT. This work represents an important step in efforts to diagnose rabies in developing countries.

Detection of rabies virus antigen by the indirect rapid immunohistochemistry test in equines and comparisons with other diagnostic techniques.

Laboratory diagnosis of rabies in equines is essential for distinguishing the disease from other sources of encephalitis. Diagnosis by conventional techniques such as a direct fluorescent antibody test (dFAT) or viral isolation in mice or cell culture can be difficult, and the application of molecular biological methods may be necessary. We performed an indirect rapid immunohistochemistry test (iRIT) for the detection of the rabies virus (RABV) antigen in the central nervous system (CNS) of equines and compared the results with those of other diagnostic techniques. We reviewed result records from the Rabies Diagnosis Laboratory at Instituto Pasteur, São Paulo, Brazil, of 174 samples of equine CNS from July 2014 to June 2016, which were investigated by dFAT, rabies tissue culture infection test (RTCIT), mouse inoculation test (MIT) and reverse transcription-polymerase chain reaction (RT-PCR) followed by genetic sequencing. These samples, 29 presented divergent results among techniques and were selected for the performed in the iRIT. The detected positivity rate was 4/29 (14%) by dFAT, 5/28 (18%) by RTCIT, 10/29 (35%) by MIT and 26/27 (96%) by RT-PCR. We analysed 29 samples through imprints of the cortex, hippocampus, cerebellum and brainstem in slides fixed in 10% buffered formaldehyde. Eighteen samples were identified as positive (62%) by iRIT assay, representing a greater number of positive cases than that detected by dFAT, MIT and RTCIT but not by RT-PCR. Among the brain regions, the brainstem presented the highest positivity (78%), followed by the hippocampus (69%), cerebellum (67%) and cortex (67%). Our results provide evidence that iRIT can contribute to a rapid diagnosis of rabies in equines and that complementary tests should be used to improve diagnostic accuracy in this species.

Performance evaluation of the polyclonal anti-rabies virus ribonucleoprotein IgG antibodies produced in-house for use in direct fluorescent antibody test.

Fluorescein isothiocyanate (FITC) labelled anti-rabies virus ribonucleoprotein (RNP) antibodies can be used as immunoreagents in direct fluorescent antibody testing (dFAT) for rabies diagnoses. While in-house products are occasionally used by laboratories, most conjugates are commercial reagents. Commercial anti-RNP antibodies are only available for research purposes in Brazil, however, which contributes to the increasing use of in-house produced antibodies. Considering that conjugate quality may influence the results obtained during rabies diagnosis, we sought to analyze the performance requirements of in-house produced polyclonal anti-RNP IgG-FITC for application in dFAT. To that end, their reproducibility, diagnostic sensitivity, and specificity were evaluated. The titer of polyclonal anti-RNP IgG-FITC was initially determined and evaluated by dFAT, using central nervous system (CNS) samples of different animal species (dogs, cats, bovines, equines, bats, and non-human primates). As our main result, the polyclonal anti-RNP IgG-FITC reached a titer of 1:30/1:40 in dFAT, with 100% of diagnostic sensitivity and specificity. In terms of reproducibility, the antibodies, regardless the production batch, presented the same performances. In conclusion, the in-house produced polyclonal anti-RNP IgG-FITC proved suitable for rabies virus antigen detection by dFAT.

Evaluation of polyclonal anti-RNP IgG antibody for rabies diagnosis by indirect rapid immunohistochemistry test.

Rabies still represents a major public health threat and estimated to cause 60,000 human deaths annually, particularly in developing countries. Thus, adequate surveillance based on rapid and reliable rabies diagnosis for both humans and animals is essential. The WHO and OIE recommended gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFAT). However, dFAT is expensive and requires a high level of expertise. As an alternative, the rapid immunohistochemistry technique is a promise to be a simple and cost effective diagnostic tool for rabies, and can be performed on field conditions prevalent in developing countries. However, no validated commercial conjugate antibody for rabies is available to meet the laboratory demand. Here, we evaluated the polyclonal anti-rabies virus ribonucleoprotein (RNP) IgG antibody for Rabies lyssavirus (RABV) detection by indirect rapid immunohistochemistry test (iRIT). We tested polyclonal anti-RNP IgG antibody against a batch of 100 brain specimens representing a wide phylogenetic origin in the State of São Paulo, Brazil. The purified IgG obtained 100% of diagnostic specificity and sensibility for RABV antigen detection in iRIT compared with the gold standard dFAT. In conclusion, our results demonstrate that the polyclonal anti-RNP IgG antibody may be used as a diagnostic reagent for rabies using iRIT, with the expectation of increase in availability and cost reduction of the epidemiological surveillance for developing countries.

Evaluation of polyclonal anti-RNP IgG antibody for rabies diagnosis by indirect rapid immunohistochemistry test

Rabies still represents a major public health threat and estimated to cause 60,000 human deaths annually, particularly in developing countries. Thus, adequate surveillance based on rapid and reliable rabies diagnosis for both humans and animals is essential. The WHO and OIE recommended gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFAT). However, dFAT is expensive and requires a high level of expertise. As an alternative, the rapid immunohistochemistry technique is a promise to be a simple and cost effective diagnostic tool for rabies, and can be performed on field conditions prevalent in developing countries. However, no validated commercial conjugate antibody for rabies is available to meet the laboratory demand. Here, we evaluated the polyclonal anti-rabies virus ribonucleoprotein (RNP) IgG antibody for Rabies lyssavirus (RABV) detection by indirect rapid immunohistochemistry test (iRIT). We tested polyclonal anti-RNP IgG antibody against a batch of 100 brain specimens representing a wide phylogenetic origin in the State of São Paulo, Brazil. The purified IgG obtained 100% of diagnostic specificity and sensibility for RABV antigen detection in iRIT compared with the gold standard dFAT. In conclusion, our results demonstrate that the polyclonal anti-RNP IgG antibody may be used as a diagnostic reagent for rabies using iRIT, with the expectation of increase in availability and cost reduction of the epidemiological surveillance for developing countries.

Evaluation of polyclonal anti-RNP IgG antibody for rabies diagnosis by indirect rapid immunohistochemistry test

Rabies still represents a major public health threat and estimated to cause 60,000 human deaths annually, particularly in developing countries. Thus, adequate surveillance based on rapid and reliable rabies diagnosis for both humans and animals is essential. The WHO and OIE recommended gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFAT). However, dFAT is expensive and requires a high level of expertise. As an alternative, the rapid immunohistochemistry technique is a promise to be a simple and cost effective diagnostic tool for rabies, and can be performed on field conditions prevalent in developing countries. However, no validated commercial conjugate antibody for rabies is available to meet the laboratory demand. Here, we evaluated the polyclonal anti-rabies virus ribonucleoprotein (RNP) IgG antibody for Rabies lyssavirus (RABV) detection by indirect rapid immunohistochemistry test (iRIT). We tested polyclonal anti-RNP IgG antibody against a batch of 100 brain specimens representing a wide phylogenetic origin in the State of São Paulo, Brazil. The purified IgG obtained 100% of diagnostic specificity and sensibility for RABV antigen detection in iRIT compared with the gold standard dFAT. In conclusion, our results demonstrate that the polyclonal anti-RNP IgG antibody may be used as a diagnostic reagent for rabies using iRIT, with the expectation of increase in availability and cost reduction of the epidemiological surveillance for developing countries.