Influência das condições de cultivo na produção de antígenos recombinantes de Leishmania i. chagasi utilizando Escherichia coli M15 cultivada em incubador rotativo e biorreator

Escherichia coli has been one of the most widely used hosts in recombinant protein production, in both laboratory and industrial scale since the advent of recombinant DNA technology. Despite the substantial progress of studies on the molecular biology and immunology of infections, there is currently no medication-based prophylaxis capable of preventing leishmaniasis. As such, there is a great need to identify specific antigens for the development of vaccines and diagnostic kits against visceral leishmaniasis. Thus, the primary goal of the present study is to assess the influence of cultivation conditions on the production of Leishmania chagasi antigens, carried out in a rotating incubator and bioreactor. To that end, several assays were conducted to evaluate the kinetic behavior of antigens (648, 503) of Leishmania. i. chagasi in two different compositions of media (2xTY, TB), with and without an inducer. In order to improve expression, assays were performed in a benchtop bioreactor using the best conditions obtained in a rotating incubator, in addition to assessing the influence of stirring speed. Results show that high complexity of the cultivation medium favored kinetic growth of clones (648, 503). However, in assays submitted to induction by IPTG, this elevated complexity did not promote the expression of recombinant proteins. Expression of antigens 648 and 503 exhibited behavior associated with growth and, in terms of location, proteins 648 and 503 are intracellularly stored. Lactose may be the most adequate inducer in protein expression, when considering factors, cost, toxicity and stability. Elevated stirring may increase cell growth in clone 53, although it may not result in high concentrations for the protein of interest. On the other hand, positive results were obtained for all recombinant clones (648, 503) tested, confirmed by the electrophoretic profile

Associação entre polimorfismos em enzimas de reparo de DNA e ocorrência de meningite bacteriana.

Despite advances in vaccine development and therapy, bacterial meningitis (BM) remains a major cause of death and long-term neurological disabilities. As part of the host inflammatory response to the invading pathogen, factors such as reactive oxygen species are generated, which may damage DNA and trigger the overactivation of DNA repair mechanisms. It is conceivable that the individual susceptibility and outcome of BM may be in part determined by non synonymous polymorphisms that may alter the function of crucial BER DNA repair enzymes as PARP-1, OGG-1 and APE-1. These enzymes, in addition to their important DNA repair function, also perform role of inflammatory regulators. In this work was investigated the non synonymous SNPs APE-1 Asn148Glu, OGG-1 Ser326Cys,PARP-1 Val762Ala, PARP-1 Pro882Leu and PARP-1 Cys908Tyr in patients with bacterial meningitis (BM), chronic meningitis (CM), aseptic meningitis (AM) and not infected (controls). As results we found increased frequency of variant alleles of PARP-1 Val762Ala (P = 0.005) and APE-1 Asn148Glu (P=0.018) in BM patients, APE-1 Asn148Glu in AM patients (P = 0.012) and decrease in the frequency of the variant allele OGG-1 Ser326Cys in patients with CM (P = 0.013), regarding the allelic frequencies in the controls. A major incidence of individuals heterozygous and/ or polymorphic homozygous in BM for PARP-1 Val762Ala (P= 0.0399, OD 4.2, 95% IC 1.213 -14.545) and PARP-1 Val762Ala/ APE-1 Asn148Glu (P = 0.0238, OD 11.111, 95% IC 1.274 - 96.914) was observed related to what was expected in a not infected population. It was also observed a major incidence of combined SNPs in the BM patients compared with the control group (P=0.0281), giving evidences that SNPs can cause some susceptibility to the disease. This combined effect of SNPs seems to regulate the principal cytokines and other factors related to BM inflammatory response and point the importance of DNA repair not only to repair activity when DNA is damaged, but to others essential functions to human organism balance.

Desenvolvimento de sistemas farmacêuticos emulsionados para veiculação gênica

Broadly speaking, the concept of gene therapy involves the transfer of a genetic material into a cell, tissue, or organ in order to cure a disease or at least improve the clinical status of a patient. Making it simple, gene therapy consists in the insertion of functional genes into cells containing defective genes by substituting, complementing or inhibiting them. The achievement of a foreigner DNA expression into a population of cells requires its transfer to the target. Therefore, it is a key issue to create systems able to transfer and protect the DNA until it reaches the target, the vectors. The disadvantages related to the use of viral vectors have encouraged efforts to develop emulsions as non-viral vectors. In fact, they are easily produced, present controllable stability and enable transfection. The aim of this work was to develop an emulsion for gene therapy and evaluate its ability to compact nucleic acids by the development of a complex with the plasmid pIRES2-EGFP. The first step was to determine the Hydrophilic Lipophilic Balance (HLB) of the Captex® 355 (oily internal phase of the emulsion) through long and short term stability assays. Based on the results, emulsions composed of Captex® 355, Tween 20® and Span 60® with 10.7 HLB were produced by three different methods: phase inversion, spontaneous emulsification and sonication. The results showed that the lowest diameter and best stability of the emulsions were achieved by the sonication method. The cationic emulsions were made by adding DOTAP to the basic emulsion. Its association with pIRES2-EGFP was evaluated by electrophoresis. Several rates of emulsion and DNA were evaluated and the results showed that 100% of the complex was formed when the rate DOTAP/DNA(nmol/µg) was 130. In conclusion, the overall results show the ability of the proposed emulsion to compact pIRES2-EGFP, which is a requirement to a successful transfection. Therefore, such formulation may be considered a promising candidate for gene therapy

Estudo da detecção do DNA do papiloma vírus humano (HPV) e da expressão imuno-histoquímica de proteína do ciclo celular no carcinoma epidermóide oral

Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral cavity and human papillomavirus (HPV) may have an important role in its development. The aim of this experiment was to investigate the HPV DNA and viral types in 90 cases of OSCC. Moreover, a comparative analysis between the cases of OSSC with and without HPV DNA was performed by using cell cycle markers p21 and pRb in order to detect a possible correlation of these proteins and HPV infection. DNA was extracted from paraffin embedded tissue and amplified by PCR (polymerase chain reaction) with primers PCO3+ e PCO4+ for a fragment of human β-globin gene. After this procedure, PCR for HPV DNA detection was realized using a pair of generic primers GP5+ e GP6+. Immunohistochemical study was performed by streptoavidin-biotin technique and antibodies against p21 and pRb proteins were employed. Eighty-eight cases were positive for human β-globin gene and HPV DNA was found in 26 (29.5%) of then. It could not be detected significant correlation between HPV and age, sex and anatomical sites of the lesion. The most prevalent viral type was HPV 18 (80.8%). Regarding the immunohistochemical analysis, it was detected significant association between HPV presence and pRb immunoexpression (p=0,044), nevertheless, the same was not observed in relation to p21 protein (p =0,416). It can be concluded that the low detection of HPV DNA in OSCC by the present experiment suggests a possible role of the virus in the development and progression in just a subset of this disease

Estimação de parâmetros genéticos para resistência à infecção por IMNV em camarões Litopenaeus vannamei por meio de análise de sobrevivência

The main specie of marine shrimp raised at Brazil and in the world is Litopenaeus vannamei, which had arrived in Brazil in the `80s. However, the entry of infectious myonecrosis virus (IMNV), causing the infectious myonecrosis disease in marine shrimps, brought economic losses to the national shrimp farming, with up to 70% of mortality in the shrimp production. In this way, the objective was to evaluate the survival of shrimps Litopenaeus vannamei infected with IMNV using the non parametric estimator of Kaplan-Meier and a model of frailty for grouped data. It were conducted three tests of viral challenges lasting 20 days each, at different periods of the year, keeping the parameters of pH, temperature, oxygen and ammonia monitored daily. It was evaluated 60 full-sib families of L. vannamei infected by IMNV in each viral challenge. The confirmation of the infection by IMNV was performed using the technique of PCR in real time through Sybr Green dye. Using the Kaplan-Meier estimator it was possible to detect significant differences (p <0.0001) between the survival curves of families and tanks and also in the joint analysis between viral challenges. It were estimated in each challenge, genetic parameters such as genetic value of family, it`s respective rate risk (frailty), and heritability in the logarithmic scale through the frailty model for grouped data. The heritability estimates were respectively 0.59; 0.36; and 0.59 in the viral challenges 1; 2; and 3, and it was also possible to identify families that have lower and higher rates of risk for the disease. These results can be used for selecting families more resistant to the IMNV infection and to include characteristic of disease resistance in L. vannamei into the genetic improvement programs

Micropartículas de quitosana estruturadas com aerosil®: estabilidade, adsorção, encapsulação e liberação de substâncias ativas

Biodegradable microspheres used as controlled release systems are important in pharmaceutics. Chitosan biopolymer represents an attractive biomaterial alternative because of its physicochemical and biological characteristics. Chitosan microspheres are expected to become promising carrier systems for drug and vaccine delivery, especially for non-invasive ways oral, mucosal and transdermal routes. Controlling the swelling rate and swelling capacity of the hydrogel and improving the fragile nature of microspheres under acidic conditions are the key challenges that need to be overcomed in order to enable the exploration of the full pharmaceutical potential use of these microparticles. Many studies have focused on the modification of chitosan microsphere structures with cross-linkers, various polymers blends and new organic-inorganic hybrid systems in order to obtain improved properties. In this work, microspheres made of chitosan and nanosized hydrophobic silica (Aerosil R972) were produced by a method consisting of two steps. First, a preparation of a macroscopically homogeneous chitosan-hydrophobic silica dispersion was prepared followed by spray drying. FTIR spectroscopy, X-ray powder diffraction, differential scanning calorimetry, thermal gravimetric analysis, scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (TEM) were used to characterize the microspheres. Also, the were conducted acid stability, moisture sorption capacity, release properties and biological assays. The chitosan-hydrophobic silica composite microspheres showed improved thermal degradation, lower water affinity, better acid stability and ability to retard rifampicin and propranolol hydrochloride (drug models) release under simulated physiological conditions. In vitro biocompatibility studies indicated low cytotoxicity and low capacity to activate cell production of the pro-inflammatory mediator nitric oxide. The results show here encourage further studies on the use of the new chitosan-hydrophobic silica composite microspheres as drug carrier systems via oral or nasal routes.

Co-circulação dos vírus dengue tipo 1, 2 e 4 nos Estados do Rio Grande do Norte e Paraíba, 2013

Dengue is a viral disease transmitted by female mosquitoes from genus Aedes, the principal urban vector is Aedes aegypti. Actually dengue has caused, in global scale, substantial morbidity and mortality. Four serotypes (antigenically distinct) are known: DENV-1, DENV-2, DENV-3 and DENV-4. The objective of this study was described the epidemiological profile dengue in the states of Rio Grande do Norte (RN) and Paraíba (PB), 2013. For that, suspected cases of dengue were studied, received for Laboratory of Molecular Biology of infectious disease and cancer (LADIC-UFRN) from different Health Units from RN and PB between January and December of 2013. The viral RNA was obtained from serum samples of patient from health units from RN and PB. It were studied 478 suspected cases of dengue , 252 (52,7%) from Rio Grande do Norte and 226 (47,3%) from Paraíba, showeds a global rate of infection global prevalence of 29,7% (142/478). The co-circulation of three serotypes was observed: DENV-1 (9,8% [14/142]), DENV-2 (3,5% [5/142]) and DENV-4 (86,7% [123/142]). People between 21-30 years old were the most affected by the disease during all the period of the study, representing 63,7% of the cases in both states. The genus most affected was female, representing 63,3% of cases in both states. Pau dos Ferros, Rio Grande do Norte, had the highest circulation of disease, with 8,2% (8/97) of cases. In Paraíba, the city most affected was João Pessoa, with (80% (36/45) of cases. The months with the biggest viral circulation in RN and PB were March and August, respectively. These results are very important to understanding the dengue viral activity in RN and PB, providing data that can guide control actions of this disease in support to local control programs

Estudo in silico das interações da protease NS3-NS2B de DENV-2 com o inibidor peptídico Bz-nKRR-H com finalidades terapêuticas

Dengue virus is an important patogen that causes Dengue desease in all world, and belongs to Flavivirus gender. The virus consists of enveloped RNA with a single strand positive sense, 11Kb genome. The RNA is translated into a polyprotein precursor, wich is cleaved into 3 structural proteins (C, prM e E) and 7 non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B e NS5). The NS3 is a multifunctional protein, that besides to promote the polyprotein precursor cleavage, also have NTPase, helicase and RTPase activity. The NS3 needs a hydrophilic segment of 40 residues from the transmembrane NS2B protein (who acts like cofator) to realize this functions. Actually, there's no vacines available on the market, and the treatment are just symptomatic. The tetrapeptide inhibitor Bz-Nle-Lys-Arg-Arg-H (Ki de 5,8-7,0 M) was showed as a potent inhibitor μ for NS3prot in Dengue virus. That is a inteligent alternative to treat the dengue desease. The present work aimed analyse the interactions of the ligand bounded to the activity site to provid a clear and depth vision of that interaction. For this purpouse, it was conducted an in silico study, by using quantum mechanical calculations based on Density Functional Theory (DFT), with Generalized Gradient approximation (GGA) to describe the effects of exchange and correlation. The interaction energy of each amino acid belonging to the binding site to the ligand was calculated the using the method of molecular fragmentation with conjugated caps (MFCC). Besides energy, we calculated the distances, types of molecular interactions and atomic groups involved. The theoretical models used were satisfactory and show a more accurate description when the dielectric constant = 20 ε and 80 was used. The results demonstrate that the interaction energy of the system reached convergence at 13.5 A. Within a radius of 13,5A the most important residues were identified. Met49, Met84 and Asp81 perform interactions of hydrogen with the ligant. The Asp79 and Asp75 residues present high energy of attraction. Arg54, Arg85 and Lys 131 perform hydrogen interactions with the ligand, however, appear in BIRD graph having high repulsion energy with the inhibitor. The data also emphasizes the importance of residue Tyr161 and the involvement of the catalytic triad composed by Asp75, His51 and Ser135

Validação do conteúdo do diagnóstico de enfermagem proteção ineficaz para pacientes submetidos à hemodiálise

Methodological study, in order to validate the content of the nursing diagnosis protection ineffective in patients undergoing hemodialysis. The research took place in two stages, namely: concept analysis and content analysis of the instrument by experts. T he first step was operationalized through an integrative review of the databaes LILACS, CINAHL, PubMed, Scopus and Cochrane, with the key words protection and hemodialysis, in October and November 2013. The sample consisted of 32 articles, which were analyz ed by a c areful reading to identify the sections that correspond ed to the defining attributes , antecedents and consequences of protection in patients undergoing hemodialysis. T he interpretation for the diagnosis of effective protection was made by transpos ing the components of the diagnosis (definition, defining characteristics and related factors) to the denial form . In the second stage, we elaborated an instrument with the components of the nursing diagnostic s studied and proceeded to the analysis conduct ed in April 2014 by 22 specialists in nephrology and in the terminology of the NANDA International, selected by means of th e Lattes Platform . We used the binomial test to assess the proportion of experts who rated each item as appropriate, considering a si gnificance level of 5%. The project was approved by the Ethics Committee of the institution responsible for the research, an opinion on num b er 387 837 and CAAE 18486413.0.0000.5537. The results show that the proposal for the nursing diagnosis of ineffectiv e protection in patients undergoing hemodialysis is: definition - the same as that presented in the NANDA International Taxonomy II, location - domain safety / protection and class injury. Related factors are: Absence of routine vaccines; Non - adherence to care related to vascular access; Non - adherence to infection control measures; Non - adherence to prescribed diet; Non - adherence to drug therapy; Presence of comorbidities; Drug abuse; Immune disorders; Extremes of age; Abnormal blood profiles; Drugs that red uce immunity; and side effects and adverse treatment - related. The defining characteristics are: Presence of invading the bloodstream; Nutritional disorders; Increase in the number of hospitalizations; Uncontrolled dry weight; Infected vascular access; Vasc ular inadequate access; Increased blood pressure; fever; Bleeding disorder; Disability immunity; fatigue; weakness; itching; and maladaptive response to stress. It follows that the identification of the defining attributes, antecedents and consequences inc reased the wealth of vocabulary, allowing the construction of theoretical and empirical definitions for a broader understanding of the concept protection. Furthermore, the study contributed to the enrichment of nursing specific body of knowledge, as well a s in the direction of nursing care for patients undergoing hemodialysis.

Spondias tuberosa Arr. (UMBU): estudo fitoquímico e avaliação do potencial anti-inflamatório

Spondias tuberosa Arruda (Anacardiaceae) is a fruitful tree popularly known as umbuzeiro, tapereba or umbu. It is a native and endemic species from Brazil, widespread in Brazilian Northeast. The species is important in folk medicine of the semi-arid Northeast, where it is mainly used to treat various inflammatory conditions, digestive problems as well as viral and bacterial infections. However, despite the common use in folk medicine, there are scarce pharmacological and phytochemicals studies that afford scientific evidence to its popular use. Therefore, this study aimed to characterize the chemical markers in S. tuberosa leaves extract, obtained by maceration ethanol:water (70:30, [v/v]), and evaluate its anti-inflammatory potential in vivo. The phytochemical profile in TLC analysis suggested the occurence of the flavonoids rutin and isoquercitrin. HPLC analysis enabled us to confirm the presence of flavonoids and also, were detected the phenolic acids, chlorogenic acid and caffeic acid. In addition was developed and validated a HPLC method to evaluate the content of the identified compounds in S. tuberosa leaves extract according to RDC 899/2003 of ANVISA and ICH Guidelines 2005. In order to evaluate the anti-inflammatory potential of S. tuberosa leaves extract, the peritonitis and paw edema models induced by carrageenan were used, administration i.p. in mice. The results highlighted the anti-inflammatory property in vivo at 125, 250 and 500 mg/kg since a decrease in leukocyte influx to the site of inflammation, diameter of the edema and the level of myeloperoxidase were observed when compared to the drug control dexamethasone (2 mg/kg, i.p. route). Taken together, the results pointed out S. tuberosa as a potential species for developing phytotherapic derivatives in according to its popular use. With regard to the characterization markers, chlorogenic acid, caffeic acid, rutin and isoquercitrin were identified and quantified in Spondias tuberosa leaves extract so they could be used in quality control analyses of the raw material and extracts of this species.

A atuação do enfermeiro no combate à dengue e a febre chikungunya: estudo comparativo nos municípios de Parnamirim e Santa Cruz/RN

The research aimed to analyze the role of nurses in the Family Health Strategy (FHS) in the control of Dengue and Chikungunya fever in the cities of Parnamirim and Santa Cruz. It is exploratory and descriptive nature of research, with a qualitative approach, which was developed with nurses of the Family Health Strategy in the municipalities of Parnamirim and Santa Cruz. All research process followed the ethical standards laid out research in Resolution No. 466/2012 of the National Health Council about research involving human beings. Data collection was performed using a semi-structured questionnaire with open and closed questions, organized into three parts: the profile of respondents, knowledge of the disease (transmission, prevention, control) and description of practices. The actions performed by nurses are punctual performed when there is growing disease of the number of cases in order to try to reduce the number of cases. The principal methodology used for them is to talk and guidelines resulting from campaigns and home visits, using leaflets and social networks. Regarding knowledge of nurses in the city of Santa Cruz on Dengue Fever and Chikungunya, is clearly some gaps. Nursing professionals know recognize a suspected case of dengue, but get confused when trying to explain to the Chikungunya fever, exhibiting the same symptoms of dengue. Although everybody had participated in a training on Chikungunya Fever and Dengue, a very limited knowledge of nurses regarding the clinical management. It is suggested that further studies are developed on the ground a few nurses have accepted to participate in the study, so that we can identify strategies, interventions, activities and nursing actions that are consistent with the reality of working in favor of coping Aedes epidemics aegypti that have positive impact on reducing the infestation rates and may be suitable and applied in other regions.

A atuação do enfermeiro no combate à dengue e a febre chikungunya: estudo comparativo nos municípios de Parnamirim e Santa Cruz/RN

The research aimed to analyze the role of nurses in the Family Health Strategy (FHS) in the control of Dengue and Chikungunya fever in the cities of Parnamirim and Santa Cruz. It is exploratory and descriptive nature of research, with a qualitative approach, which was developed with nurses of the Family Health Strategy in the municipalities of Parnamirim and Santa Cruz. All research process followed the ethical standards laid out research in Resolution No. 466/2012 of the National Health Council about research involving human beings. Data collection was performed using a semi-structured questionnaire with open and closed questions, organized into three parts: the profile of respondents, knowledge of the disease (transmission, prevention, control) and description of practices. The actions performed by nurses are punctual performed when there is growing disease of the number of cases in order to try to reduce the number of cases. The principal methodology used for them is to talk and guidelines resulting from campaigns and home visits, using leaflets and social networks. Regarding knowledge of nurses in the city of Santa Cruz on Dengue Fever and Chikungunya, is clearly some gaps. Nursing professionals know recognize a suspected case of dengue, but get confused when trying to explain to the Chikungunya fever, exhibiting the same symptoms of dengue. Although everybody had participated in a training on Chikungunya Fever and Dengue, a very limited knowledge of nurses regarding the clinical management. It is suggested that further studies are developed on the ground a few nurses have accepted to participate in the study, so that we can identify strategies, interventions, activities and nursing actions that are consistent with the reality of working in favor of coping Aedes epidemics aegypti that have positive impact on reducing the infestation rates and may be suitable and applied in other regions.

Vacinação: o fazer da enfermagem e o saber das mães e/ou cuidadores

Observa-se na prática das unidades de saúde, ainda hoje, o pouco conhecimento das mães sobre o processo de vacinação e pouco envolvimento do enfermeiro nesse processo, principalmente na área de educação em saúde. O objetivo é identificar as ações de enfermagem na sala de vacina e descrever o conhecimento das mães/cuidadores acerca da vacinação infantil. Estudo exploratório-descritivo com abordagem quantitativa. A população foi composta por 43 mães e/ou cuidadores e 10 profissionais de enfermagem. Ocorreu em duas Unidades de Saúde da Família, na Zona Oeste de Natal/RN, em novembro e dezembro/2008. A maioria das mães (88,4%) conhece a importância de vacinar a crianças, embora não saibam quais as vacinas estão sendo dadas e para quais doenças são destinadas. A maioria dos profissionais prioriza mais a técnica do que a atividade educativa. Percebe-se, portanto, que há lacunas no saber das mães/cuidadores e nas ações de enfermagem que visem um trabalho promocional na sala de vacinação

O enfermeiro na sala de vacinação: uma análise reflexiva da prática

Objetiva-se analisar a prática do enfermeiro em sala de vacina, mediante o modelo de vigilância à saúde como referência. Trata-se da análise crítica reflexiva realizada através de observação, durante as práticas como docentes, em Unidades Básicas de Saúde. Percebeu-se que o enfermeiro responsável pela sala de vacinação desenvolve pouca atividade fixa específica, indo à sala para resolver algumas dificuldades ou na ausência de funcionários. O perfil epidemiológico e o modelo de vigilância à saúde são pouco usados, sinalizado pelo comportamento indiferente dos usuários. As atividades de vacinação devem ser desenvolvidas por uma equipe de enfermagem com participação do enfermeiro, a fim de oferecer um serviço de qualidade e atender a demanda individual e coletiva

Estudo do genoma do vírus causador da mionecrose infecciosa em camarões e desenvolvimento de métodos para detecção de polimorfismos

Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites