Avaliação da acurácia da proteína rKLO8 no diagnóstico da leishmaniose visceral canina

A leishmaniose visceral canina (LVC) representa um grave problema de saúde pública. No Brasil, a prevalência da infecção nos cães é bastante variável, podendo atingir níveis superiores a 60% em alguns surtos. O teste rápido Dual Path Platform (TRDPP®-Bio-Manguinhos), como teste de triagem, seguido por ELISA (EIE-BioManguinhos), como teste confirmatório, tornaram-se parte do protocolo de diagnóstico da LVC, credenciado no Brasil desde 2011. No entanto, o diagnóstico da LVC ainda precisa ser melhorado para alcançar uma taxa de detecção mais precisa. Recentemente, rKLO8, uma nova proteína antigênica de L. donovani do Sudão foi clonada e purificada, e mostrou alta reatividade para diagnosticar leishmaniose visceral em humanos. O presente estudo teve como objetivo avaliar a reatividade de soros de cães frente ao antígeno rKL08 e o antígeno de referência rK26, comparando ambas as proteínas, utilizadas como antígenos em testes de ELISA, com os testes DPP® e EIE, usados como testes de diagnóstico da LVC. Amostras de soros de cães de Governador Valadares, uma área endêmica para leishmaniose em Minas Gerais, Brasil, foram agrupadas da seguinte forma: (I) DPP®/EIE negativo (n = 100), (II) DPP® positivo / EIE negativo e (III) DPP® / EIE positivo (n = 100). Níveis séricos elevados de IgM e IgG para ambos os antígenos, rKLO8 e rK26, foram encontrados no grupo III (p <0,0001). Interessantemente, foram detectados níveis elevados de IgG2 e baixos níveis de IgG1 contra ambos os antígenos no grupo de cães DPP®/EIE positivo, sugerindo a ocorrência de um fenótipo predominantemente do tipo Th1 associado com infecção subclínica. O ELISA-rKLO8 (IgG) e o ELISA-rK26 (IgG) mostraram uma sensibilidade de 68% e 77%, e especificidade de 92% e 91%, respectivamente, determinado através da análise da curva ROC. Além disso, o coeficiente Kappa indicou boa concordância (0,739) entre o ELISA-rKLO8 versus o ELISA-rK26. Ainda, a combinação de antígenos rKLO8 e rK26 (rKLO8+rK26) em um mesmo teste exibiu maior sensibilidade (85%) e especificidade (93%). A análise kappa mostrou que o ELISA-rKLO8 + rK26 (IgG) teve melhor concordância com ambos os testes, DPP® e EIE, com valores de kappa igual a 0,700. Estes dados indicaram que a combinação dos antígenos rKLO8 e rK26 gera uma melhor acurácia no diagnóstico da LVC que os antígenos rKLO8 e rK26 usados em separado na detecção de IgG. Estes resultados demonstraram, pela primeira vez, a utilidade do antígeno rKLO8 no diagnóstico da LVC, e que ELISA-rKLO8, pode representar uma potencial ferramenta adicional para o diagnóstico de LVC.Canine Visceral Leishmaniasis (CVL) represents a serious public health issue. In Brazil, the prevalence of infection in dogs is quite variable and may reach levels above 60% in some outbreaks. The dual Path Platform (DPP®-Bio-Manguinhos) as quick screening test followed by ELISA (EIE-Bio-Manguinhos) as a confirmatory test became part of the diagnostic protocol of CVL, nationally accreditated in Brazil since 2011. However, CVL diagnosis still needs to be improved to achieve a more accurate detection rate. Recently, rKLO8, a new antigenic protein of Sudanese L. donovani, was cloned and purified and had high reactivity to diagnose human VL. The present study aimed to evaluate serum reactivity to rKL08 and to the reference antigen rK26, and to compare both diagnostic proteins used in ELISA with the combined DPP® and EIE as diagnostic tests of CVL. Dog sera samples from Governador Valadares, an area endemic for leishmaniasis in Minas Gerais, Brazil, were grouped in the following way: (I) DPP®/EIE negative (n = 100), (II) DPP® positive/EIE negative and (III) DPP®/EIE positive dog sera (n = 100). Enhanced serum levels of IgM and IgG to both rKLO8 and rK26 were found in group III (p<0.0001). Interestingly, high IgG2 and low IgG1 levels against both antigens were detected in DPP®/EIE positive dogs, suggesting the occurrence of a predominant Th1 phenotype associated with subclinical infection. The rKLO8-ELISA (IgG) and the rK26-ELISA (IgG) showed a sensitivity of 68% and 77% and specificity of 92% and 91%, respectively, determined by ROC curve analysis. In addition, Kappa coefficient indicated good agreement (0.739) between rKLO8-ELISA and rK26-ELISA. Moreover, the combination of rKLO8 and rK26 antigens (rKLO8+rK26) exhibited higher sensitivity (85%) and specificity (93%). Kappa analysis established that rKLO8+rK26-ELISA (IgG) had better agreement with both DPP® and EIE, with kappa values of 0.700. These data indicate that the combination of rKLO8 and rK26 antigens has better accuracy in the diagnosis of CVL than rKLO8 and rK26 used separately at detecting IgG. These results showed for the first time the usefulness of rKLO8 antigen in the diagnosis of CVL, and that rKLO8-ELISA may represent a potential additional tool for the diagnosis of CVL

Avaliação da acurácia da proteína rKLO8 no diagnóstico da leishmaniose visceral canina

Canine Visceral Leishmaniasis (CVL) represents a serious public health issue. In Brazil, the prevalence of infection in dogs is quite variable and may reach levels above 60% in some outbreaks. The dual Path Platform (DPP®-Bio-Manguinhos) as quick screening test followed by ELISA (EIE-Bio-Manguinhos) as a confirmatory test became part of the diagnostic protocol of CVL, nationally accreditated in Brazil since 2011. However, CVL diagnosis still needs to be improved to achieve a more accurate detection rate. Recently, rKLO8, a new antigenic protein of Sudanese L. donovani, was cloned and purified and had high reactivity to diagnose human VL. The present study aimed to evaluate serum reactivity to rKL08 and to the reference antigen rK26, and to compare both diagnostic proteins used in ELISA with the combined DPP® and EIE as diagnostic tests of CVL. Dog sera samples from Governador Valadares, an area endemic for leishmaniasis in Minas Gerais, Brazil, were grouped in the following way: (I) DPP®/EIE negative (n = 100), (II) DPP® positive/EIE negative and (III) DPP®/EIE positive dog sera (n = 100). Enhanced serum levels of IgM and IgG to both rKLO8 and rK26 were found in group III (p<0.0001). Interestingly, high IgG2 and low IgG1 levels against both antigens were detected in DPP®/EIE positive dogs, suggesting the occurrence of a predominant Th1 phenotype associated with subclinical infection. The rKLO8-ELISA (IgG) and the rK26-ELISA (IgG) showed a sensitivity of 68% and 77% and specificity of 92% and 91%, respectively, determined by ROC curve analysis. In addition, Kappa coefficient indicated good agreement (0.739) between rKLO8-ELISA and rK26-ELISA. Moreover, the combination of rKLO8 and rK26 antigens (rKLO8+rK26) exhibited higher sensitivity (85%) and specificity (93%). Kappa analysis established that rKLO8+rK26-ELISA (IgG) had better agreement with both DPP® and EIE, with kappa values of 0.700. These data indicate that the combination of rKLO8 and rK26 antigens has better accuracy in the diagnosis of CVL than rKLO8 and rK26 used separately at detecting IgG. These results showed for the first time the usefulness of rKLO8 antigen in the diagnosis of CVL, and that rKLO8-ELISA may represent a potential additional tool for the diagnosis of CVL.A leishmaniose visceral canina (LVC) representa um grave problema de saúde pública. No Brasil, a prevalência da infecção nos cães é bastante variável, podendo atingir níveis superiores a 60% em alguns surtos. O teste rápido Dual Path Platform (TRDPP®-Bio-Manguinhos), como teste de triagem, seguido por ELISA (EIE-BioManguinhos), como teste confirmatório, tornaram-se parte do protocolo de diagnóstico da LVC, credenciado no Brasil desde 2011. No entanto, o diagnóstico da LVC ainda precisa ser melhorado para alcançar uma taxa de detecção mais precisa. Recentemente, rKLO8, uma nova proteína antigênica de L. donovani do Sudão foi clonada e purificada, e mostrou alta reatividade para diagnosticar leishmaniose visceral em humanos. O presente estudo teve como objetivo avaliar a reatividade de soros de cães frente ao antígeno rKL08 e o antígeno de referência rK26, comparando ambas as proteínas, utilizadas como antígenos em testes de ELISA, com os testes DPP® e EIE, usados como testes de diagnóstico da LVC. Amostras de soros de cães de Governador Valadares, uma área endêmica para leishmaniose em Minas Gerais, Brasil, foram agrupadas da seguinte forma: (I) DPP®/EIE negativo (n = 100), (II) DPP® positivo / EIE negativo e (III) DPP® / EIE positivo (n = 100). Níveis séricos elevados de IgM e IgG para ambos os antígenos, rKLO8 e rK26, foram encontrados no grupo III (p <0,0001). Interessantemente, foram detectados níveis elevados de IgG2 e baixos níveis de IgG1 contra ambos os antígenos no grupo de cães DPP®/EIE positivo, sugerindo a ocorrência de um fenótipo predominantemente do tipo Th1 associado com infecção subclínica. O ELISA-rKLO8 (IgG) e o ELISA-rK26 (IgG) mostraram uma sensibilidade de 68% e 77%, e especificidade de 92% e 91%, respectivamente, determinado através da análise da curva ROC. Além disso, o coeficiente Kappa indicou boa concordância (0,739) entre o ELISA-rKLO8 versus o ELISA-rK26. Ainda, a combinação de antígenos rKLO8 e rK26 (rKLO8+rK26) em um mesmo teste exibiu maior sensibilidade (85%) e especificidade (93%). A análise kappa mostrou que o ELISA-rKLO8 + rK26 (IgG) teve melhor concordância com ambos os testes, DPP® e EIE, com valores de kappa igual a 0,700. Estes dados indicaram que a combinação dos antígenos rKLO8 e rK26 gera uma melhor acurácia no diagnóstico da LVC que os antígenos rKLO8 e rK26 usados em separado na detecção de IgG. Estes resultados demonstraram, pela primeira vez, a utilidade do antígeno rKLO8 no diagnóstico da LVC, e que ELISA-rKLO8, pode representar uma potencial ferramenta adicional para o diagnóstico de LVC

Circulación silenciosa de Leishmania infantum y Trypanosoma cruzi en perros domésticos de áreas urbanas de Sincelejo, región Caribe de Colombia

Introduction: Leishmania infantum and Trypanosoma cruzi are considered endemic zoonotic agents from rural areas of the country, however, there is a high risk of urbanization due to anthropogenic processes. For this reason, dogs have been proposed as sentinels of these zoonoses given their role as patients, hosts and/or reservoirs.&nbsp; Objective: to assess the potential circulation of Leishmania and T. cruzi parasites in canines from urban areas of Sincelejo, Sucre.&nbsp; Materials and methods: The presence of Leishmania and T. cruzi was evaluated by PCR, while identification of Leishmania was carried out by RFLP and DNA sequencing. In addition, the presence of E. canis and A. platys was evaluated as infections that can influence the clinical symptoms and health of animals. Results: Leishmania was detected in 32% and T. cruzi in 12% of the animals, 7% of the samples were positive for both parasites and L. infantum was detected in 18%. Anaplasmataceae infections were detected in 18% of the animals, co-infections by bacteria and parasites were found in 8% of the animals. Overall, 47% of the animals were infected by at least one agent, and these infections were associated with 50% of the total symptomatic animals.&nbsp; Conclusion: the silent circulation of L. infantum and T. cruzi is demonstrated in urban areas of Sincelejo, as well as co-infections of parasites with anaplasmataceae. The present study demonstrates the convenience of the use of canines in the surveillance of these zoonotic agents.Introducción: La enfermedad de Chagas y la leishmaniasis tradicionalmente se han considerado zoonosis endémicas de áreas rurales del país, sin embargo, la aparición de casos de estas enfermedades en áreas urbanas sugiere nuevos ciclos de circulación de estos parásitos. Por ello, se ha propuesto a los perros como centinelas de estos agentes zoonóticos dado su rol de hospederos accidentales o reservorios. Objetivo: evaluar la circulación silenciosa de Leishmania y T. cruzi en perros residentes de zonas urbanas de la ciudad de Sincelejo, Sucre. Materiales y métodos: 100 muestras de sangre de caninos se emplearon para amplificar la región ITS1 de Leishmania, las muestras positivas fueron utilizadas para amplificar la región conservada del minicírculo del ADN del kinetoplasto de L. infantum y para el análisis de restricción con la endonucleasa HaeIII. Por otra parte, se amplificó un fragmento del DNA satelital de T. cruzi. Adicionalmente, se evaluó la presencia de E. canis y A. platys como infecciones potencialmente modificadoras de los cuadros clínicos. Resultados: Leishmania fue detectada en 32% (32/100) y T. cruzi en 12%(12/100) de los perros estudiados. El 7% (7/100) de los perros fueron positivos a ambos parásitos. Se detectó L. infantum en 18% (18/100) de los perros. Infecciones por anaplasmatáceos fueron detectadas en 18% (18/100) de los perros, se encontró coinfecciones por bacterias y parásitos en 8% (8/100) de los perros. En general, 47% (47/100) de los animales fueron infectados por al menos un agente etiológico. Conclusión: se demuestra la circulación de L. infantum y T. cruzi en zonas urbanas de Sincelejo, así como coinfecciones de estos parásitos con anaplasmatáceos. El presente estudio demuestra la conveniencia del uso de caninos en la vigilancia de estos agentes zoonóticos

Risk of COVID-19 after natural infection or vaccinationResearch in context

Summary: Background: While vaccines have established utility against COVID-19, phase 3 efficacy studies have generally not comprehensively evaluated protection provided by previous infection or hybrid immunity (previous infection plus vaccination). Individual patient data from US government-supported harmonized vaccine trials provide an unprecedented sample population to address this issue. We characterized the protective efficacy of previous SARS-CoV-2 infection and hybrid immunity against COVID-19 early in the pandemic over three-to six-month follow-up and compared with vaccine-associated protection. Methods: In this post-hoc cross-protocol analysis of the Moderna, AstraZeneca, Janssen, and Novavax COVID-19 vaccine clinical trials, we allocated participants into four groups based on previous-infection status at enrolment and treatment: no previous infection/placebo; previous infection/placebo; no previous infection/vaccine; and previous infection/vaccine. The main outcome was RT-PCR-confirmed COVID-19 >7–15 days (per original protocols) after final study injection. We calculated crude and adjusted efficacy measures. Findings: Previous infection/placebo participants had a 92% decreased risk of future COVID-19 compared to no previous infection/placebo participants (overall hazard ratio [HR] ratio: 0.08; 95% CI: 0.05–0.13). Among single-dose Janssen participants, hybrid immunity conferred greater protection than vaccine alone (HR: 0.03; 95% CI: 0.01–0.10). Too few infections were observed to draw statistical inferences comparing hybrid immunity to vaccine alone for other trials. Vaccination, previous infection, and hybrid immunity all provided near-complete protection against severe disease. Interpretation: Previous infection, any hybrid immunity, and two-dose vaccination all provided substantial protection against symptomatic and severe COVID-19 through the early Delta period. Thus, as a surrogate for natural infection, vaccination remains the safest approach to protection. Funding: National Institutes of Health

Risk of COVID-19 after natural infection or vaccinationResearch in context

Background: While vaccines have established utility against COVID-19, phase 3 efficacy studies have generally not comprehensively evaluated protection provided by previous infection or hybrid immunity (previous infection plus vaccination). Individual patient data from US government-supported harmonized vaccine trials provide an unprecedented sample population to address this issue. We characterized the protective efficacy of previous SARS-CoV-2 infection and hybrid immunity against COVID-19 early in the pandemic over three-to six-month follow-up and compared with vaccine-associated protection. Methods: In this post-hoc cross-protocol analysis of the Moderna, AstraZeneca, Janssen, and Novavax COVID-19 vaccine clinical trials, we allocated participants into four groups based on previous-infection status at enrolment and treatment: no previous infection/placebo; previous infection/placebo; no previous infection/vaccine; and previous infection/vaccine. The main outcome was RT-PCR-confirmed COVID-19 >7–15 days (per original protocols) after final study injection. We calculated crude and adjusted efficacy measures. Findings: Previous infection/placebo participants had a 92% decreased risk of future COVID-19 compared to no previous infection/placebo participants (overall hazard ratio [HR] ratio: 0.08; 95% CI: 0.05–0.13). Among single-dose Janssen participants, hybrid immunity conferred greater protection than vaccine alone (HR: 0.03; 95% CI: 0.01–0.10). Too few infections were observed to draw statistical inferences comparing hybrid immunity to vaccine alone for other trials. Vaccination, previous infection, and hybrid immunity all provided near-complete protection against severe disease. Interpretation: Previous infection, any hybrid immunity, and two-dose vaccination all provided substantial protection against symptomatic and severe COVID-19 through the early Delta period. Thus, as a surrogate for natural infection, vaccination remains the safest approach to protection. Funding: National Institutes of Health

Global variation in postoperative mortality and complications after cancer surgery: a multicentre, prospective cohort study in 82 countries

© 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 licenseBackground: 80% of individuals with cancer will require a surgical procedure, yet little comparative data exist on early outcomes in low-income and middle-income countries (LMICs). We compared postoperative outcomes in breast, colorectal, and gastric cancer surgery in hospitals worldwide, focusing on the effect of disease stage and complications on postoperative mortality. Methods: This was a multicentre, international prospective cohort study of consecutive adult patients undergoing surgery for primary breast, colorectal, or gastric cancer requiring a skin incision done under general or neuraxial anaesthesia. The primary outcome was death or major complication within 30 days of surgery. Multilevel logistic regression determined relationships within three-level nested models of patients within hospitals and countries. Hospital-level infrastructure effects were explored with three-way mediation analyses. This study was registered with ClinicalTrials.gov, NCT03471494. Findings: Between April 1, 2018, and Jan 31, 2019, we enrolled 15 958 patients from 428 hospitals in 82 countries (high income 9106 patients, 31 countries; upper-middle income 2721 patients, 23 countries; or lower-middle income 4131 patients, 28 countries). Patients in LMICs presented with more advanced disease compared with patients in high-income countries. 30-day mortality was higher for gastric cancer in low-income or lower-middle-income countries (adjusted odds ratio 3·72, 95% CI 1·70–8·16) and for colorectal cancer in low-income or lower-middle-income countries (4·59, 2·39–8·80) and upper-middle-income countries (2·06, 1·11–3·83). No difference in 30-day mortality was seen in breast cancer. The proportion of patients who died after a major complication was greatest in low-income or lower-middle-income countries (6·15, 3·26–11·59) and upper-middle-income countries (3·89, 2·08–7·29). Postoperative death after complications was partly explained by patient factors (60%) and partly by hospital or country (40%). The absence of consistently available postoperative care facilities was associated with seven to 10 more deaths per 100 major complications in LMICs. Cancer stage alone explained little of the early variation in mortality or postoperative complications. Interpretation: Higher levels of mortality after cancer surgery in LMICs was not fully explained by later presentation of disease. The capacity to rescue patients from surgical complications is a tangible opportunity for meaningful intervention. Early death after cancer surgery might be reduced by policies focusing on strengthening perioperative care systems to detect and intervene in common complications. Funding: National Institute for Health Research Global Health Research Unit

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

© 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licenseBackground: Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide. Methods: A multimethods analysis was performed as part of the GlobalSurg 3 study—a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital. Findings: Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3·85 [95% CI 2·58–5·75]; p<0·0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63·0% vs 82·7%; OR 0·35 [0·23–0·53]; p<0·0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer. Interpretation: Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised. Funding: National Institute for Health and Care Research