Detection of glycoproteins B and H genotypes to predict the development of Cytomegalovirus disease in solid organ transplant recipients.

BACKGROUND: Our study focuses on the role that human Cytomegalovirus (CMV) genotypes play in the development of disease. OBJECTIVES: (1) To analyze the frequency of various genotype envelope proteins (gB, gH) in a group of solid organ transplant (SOT) recipients; (2) to assess their correlation with CMV disease; (3) to study the association between any of the genotypes and viral loads. STUDY DESIGN: A retrospective observational study conducted by analyzing CMV gB and gH genotypes detected with real-time polymerase chain reaction (PCR)-specific assays in 162 CMV-positive blood samples from 62 SOT recipients. Demographic, clinical, and microbiological data were recorded. RESULTS: Mixed gB genotypes were associated with viral syndrome (70%, p = .004), earlier presentation of symptoms (48.27 ± 27.03 versus 74.33 ± 47.25 days, respectively, p = .001), and higher median of the plasma viral load log10 (UI/ml) than infection with a single genotype (p = .004). Furthermore, the gB3 genotype was detected more frequently in patients who presented with asymptomatic viremia (77.27%, p < .0001). The gH1 genotype was more frequent (65%) in patients who presented with asymptomatic viremia (p = .003), and it caused infection later than gH2 or the mixed genotype (84.88 ± 48.10 versus 57.91 ± 39.18 days, respectively, p < .001). CONCLUSIONS: Patients who presented mixed gB genotypes more frequently developed clinical manifestations and earlier, higher, plasma viral loads. The detection of gB and gH genotypes by real-time PCR can provide relevant information to stratify the risk of SOT recipients to develop symptomatic infection by CMV.

Evaluation of a new rapid diagnostic test for the detection of influenza and RSV / Evaluación de un nuevo test de diagnóstico rápido para la detección de influenza y VRS

INTRODUCTION: Influenza viruses and respiratory syncytial virus (RSV) can cause an acute respiratory disease that occurs seasonally in epidemic waves. This retrospective study was conducted to evaluate the Sofia®Influenza A + B and the Sofia® RSV fluorescence immunoassays (FIAs), two novel rapid detection tests (RDTs) for influenza A and B and RSV. METHODS: Two hundred and nine breath samples were selected from patients with respiratory symptoms determined to be positive/negative for influenza A, influenza B or RSV using one of the reference diagnostic techniques, cell culture and/or RT-PCR (Simplexa™Flu A/B & RSV). The Sofia Influenza A + B FIA was tested on 123 samples (63 from children and 60 from adults) and the Sofia RSV FIA was tested on 86 pediatric samples. Sensitivity and specificity values of both assays were calculated assuming the reference techniques as the gold standard. RESULTS: Sensitivity and specificity values for the Sofia Influenza A + B FIA were 73.1% and 97.8%, respectively. Sensitivity and specificity values for the Sofia RSV FIA were 87.5% and 86.7%, respectively. CONCLUSION: The sensitivity results obtained for the two assays were considerably higher than those reported for other RDTs. In conclusion, the Sofia Influenza A + B and the Sofia RSV FIAs are appropriate tools for the rapid diagnosis of these viruses

INTRODUCCIÓN: Los virus influenza A y B y el virus respiratorio sincitial (VRS) causan infecciones respiratorias agudas de forma estacional en olas epidémicas. Este estudio restrospectivo se llevó a cabo para evaluar dos nuevos tests de diagnóstico rápido para detectar los virus influenza A y B y VRS: Sofia® Influenza A + B y Sofia® RSV Fluorescence Immunoassays (FIAs). MÉTODOS: Se seleccionaron 209 muestras respiratorias procedentes de pacientes con síntomas respiratorios en las que se había establecido el diagnóstico de presencia o ausencia de influenza A, influenza B o VRS mediante las técnicas de referencia: cultivo celular o RT-PCR PCR (Simplexa™Flu A/B & RSV). Sofia Influenza A + B se realizó en 123 muestras (63 de niños y 60 de adultos) y Sofia RSV en 86 muestras pediátricas. Se calcularon los valores de sensibilidad y especificidad de ambos test tomando las técnicas de referencia como gold standard. RESULTADOS: Los valores de sensibilidad y especificidad de Sofia Influenza A + B fueron 73,1 y 97,8%. Los valores de sensibilidad y especificidad de Sofia RSV fueron 87,5 y 86,7%. CONCLUSIONES: Los valores de sensibilidad obtenidos para ambos tests son mayores que los descritos para otras técnicas de diagnóstico rápido. Por lo tanto, Sofia Influenza A + B FIA y Sofia RSV FIA son test adecuados para el diagnóstico rápido de influenza y VRS

A prospective study to assess the diagnostic performance of the Sofia(®) Immunoassay for Influenza and RSV detection.

BACKGROUND: Respiratory viruses RSV and influenza A and B viruses are responsible for important disease outbreaks during the winter season in temperate climate regions. Rapid diagnostic tests (RDTs) are assays designed to yield a rapid diagnosis, which facilitates patient management. The Sofia Influenza A+B Fluorescence Immunoassay and Sofia RSV Fluorescence Immunoassay are RDTs for Influenza and RSV detection that employ a new technology to enhance their sensitivity. OBJECTIVES: Sensitivity, specificity and positive and negative predictive values of the assays were calculated compared with the reference diagnostic method: real-time RT-PCR. STUDY DESIGN: A prospective evaluation was carried out on 1065 respiratory samples for Sofia Influenza A+B FIA and on 261 samples for Sofia RSV FIA from November 2013 to April 2014. RESULTS: The sensitivities of the Sofia Influenza A+B FIA for influenza A and influenza B detection were, respectively, 75.3% (244/324) and 50.0% (8/16). The sensitivity of the Sofia RSV FIA was 92.1% (128/139). There were no differences in Sofia FIA performance depending on the virus subtype. CONCLUSIONS: The results showed high sensitivity and specificity values for influenza A and RSV detection, but values were lower for influenza B. More information is needed regarding the performance for influenza B given the small number of positive samples assessed.

Evaluation of a new rapid diagnostic test for the detection of influenza and RSV.

INTRODUCTION: Influenza viruses and respiratory syncytial virus (RSV) can cause an acute respiratory disease that occurs seasonally in epidemic waves. This retrospective study was conducted to evaluate the Sofia(®) Influenza A+B and the Sofia(®) RSV fluorescence immunoassays (FIAs), two novel rapid detection tests (RDTs) for influenza A and B and RSV. METHODS: Two hundred and nine breath samples were selected from patients with respiratory symptoms determined to be positive/negative for influenza A, influenza B or RSV using one of the reference diagnostic techniques, cell culture and/or RT-PCR (Simplexa™Flu A/B & RSV). The Sofia Influenza A+B FIA was tested on 123 samples (63 from children and 60 from adults) and the Sofia RSV FIA was tested on 86 pediatric samples. Sensitivity and specificity values of both assays were calculated assuming the reference techniques as the gold standard. RESULTS: Sensitivity and specificity values for the Sofia Influenza A+B FIA were 73.1% and 97.8%, respectively. Sensitivity and specificity values for the Sofia RSV FIA were 87.5% and 86.7%, respectively. CONCLUSION: The sensitivity results obtained for the two assays were considerably higher than those reported for other RDTs. In conclusion, the Sofia Influenza A+B and the Sofia RSV FIAs are appropriate tools for the rapid diagnosis of these viruses.

Cytomegalovirus Genotype Distribution Among Congenitally and Postnatally Infected Patients: Association of Particular Glycoprotein (g)B and gN Types With Symptomatic Disease.

Background. Human cytomegalovirus is a leading cause of congenital infection, and there are limited data on prognosis markers in disease development. We aimed to study 3 virology targets (glycoprotein [g]B, gN, and UL144) to assess their correlation with congenital infection and various organ system involvement. Methods. Forty-eight congenital cases and 58 postnatally infected children were included (2003-2014). Genotyping for the 3 targets and distribution among the cohorts were investigated, and the relationship between the gB, gN, and UL144 types with clinical manifestations in congenital infection was also studied. Results. All of the genotypes were similarly represented among cohorts, and the most prevalent were the UL144B, gB1, and gN1 genotypes. The gB2 genotype was associated with abnormal image findings by ultrasound and/or magnetic resonance in congenital infection (odds ratio [OR], 6.2; 95% confidence interval [CI], 1.1-34.3; P = .036); the gN1 genotype was associated with an elevated risk of developing neurological disorders (OR, 7.0; 95% CI, 1.1-45.9; P = .043). Both gN1 and gB2 were independent factors for symptomatic infection. Statistical analyses showed no association between any UL144 genotype and disease severity. Conclusions. All of the genotypes can be involved in congenital infection, although the gB2 and gN1 genotypes might be associated with a more serious illness.

Benzothiadiazine dioxide human cytomegalovirus inhibitors: synthesis and antiviral evaluation of main heterocycle modified derivatives.

The benzothiadiazine dioxide derivatives are potent non-nucleoside human cytomegalovirus (HCMV) inhibitors. As part of our comprehensive structure-activity relationship (SAR) study of these compounds, we have now proposed structural modifications on the heterocyclic moiety both on the number and the nature of the fused heterocycle and on the kind of heteroatoms present on it. Synthesis of these new compounds (benzyl derivatives of thiadiazines, thienothiadiazines, benzothienothiadiazines and quinazolines) and the antiviral evaluation against HCMV has been performed. SAR investigation on this class of compounds has defined the structural requirements for potency and toxicity. They have revealed two important clues: i) a fused ring to the thiadiazine framework is necessary to maintain the anti-HCMV action, and ii) the sulfamido moiety in the main heterocycle is crucial to avoid cytotoxicity.

Benzothiadiazine dioxides (BTD) derivatives as non-nucleoside human cytomegalovirus (HCMV) inhibitors. study of structural requirements for biological activity.

Two new series of BTD derivatives have been synthesised allowing to explore the steric requirements for their biological activity. The N3-alkylBTD compounds have shown antiviral activity in the same order or lower than previously prepared compounds. However, the cytotoxicity values observed prevent this new series of BTD derivatives from its potential therapeutic application. Concerning BTD derivatives with the modified linker attached to N1 position, we have obtained new non-nucleoside anti-HCMV derivatives. The activity against HCMV is shown at concentrations that were 10-fold lower than the concentration that was toxic for the host cells, which confirm that these derivatives show a specific antiviral effect against HCMV. SAR conclusions derived from these last compounds have provided new knowledge about the structural requirements of BTD showing certain positions that could be modified for enhancing the anti-HCMV action.