Projeto Quantizika Humano-UFG / Human Quantizika Project-UFG

O projeto QUANTIZIKA HUMANO, desenvolvido por docentes de várias unidades acadêmicas da Universidade Federal de Goiás (UFG), teve início em 2016, quando um grupo de pesquisadores composto por professores das áreas de biologia molecular e genética, epidemiologista e médico sanitarista, selecionaram pacientes que apresentavam sintomas semelhantes a dengue, inicialmente com a finalidade de rastrear aqueles que pudessem apresentar infecção pelo vírus da Zika. Durante o rastreamento molecular, e por meio de técnica própria desenvolvida na pesquisa, foram identificados também casos de infecção pelo vírus Mayaro (FERNANDES, M. 2019).

The QUANTIZIKA HUMANO project, developed by professors from several academic units of the Federal University of Goiás (UFG), began in 2016, when a group of researchers composed of professors from the fields of molecular biology and genetics, epidemiologist and sanitary doctor, selected patients with symptoms similar to dengue, initially with the purpose of tracking those who could present zika virus infection. During molecular screening, and through its own technique developed in the research, cases of Mayaro virus infection were also identified (FERNANDES, M. 2019).

Momento de considerar otras arbovirosis luego del virus mayaro / Time to Consider Other Arbovirus Diseases after mayaro Virus

Introducción: En los últimos años, debido a los movimientos migratorios, se ha desarrollado una expansión de nuevas enfermedades, como chikungunya, zika, oropuche y mayaro. Caso clínico: Paciente que manifestaba síntomas de fiebre, cefalea y artralgias persistente. Después de un arduo estudio y eliminación de otras patologías se llega al diagnóstico de virus mayaro. El paciente residía en una zona nororiental del Perú. Se brindó tratamiento de soporte junto con hidratación, paracetamol 500 mg cada 8 horas y se indicó cita diaria para evaluación. El paciente evolucionó favorablemente a los pocos días. Conclusiones: La vigilancia, las pruebas y el control vectorial siguen siendo claves para prevenir la propagación de este tipo de virus. La posibilidad de que el virus mayaro se urbanice aún más. Se debe tener siempre en cuenta el diagnóstico diferencial de virus mayaro(AU)

Introduction: In recent years, due to migratory movements, an expansion of new diseases has developed, such as chikungunya, zika, oropuche and mayaro. Clinical case: Patient with the following symptoms: fever, headache and persistent arthralgia. After an arduous study and ruling out other possible diseases, we diagnose mayaro virus. The patient resided in a northeastern part of Peru. Supportive treatment was provided along with hydration; paracetamol 500 mg every 8 hours and daily appointment for evaluation was indicated. The patient evolved favorably within a few days. Conclusions: Surveillance, testing and vector control are still key to monitoring and preventing the spread of this type of virus. The possibility of mayaro virus becoming more urbanized is worthy of attention. The differential diagnosis of mayaro virus should always be considered(AU)

Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses

ABSTRACT We describe a sensitive method for simultaneous detection of Oropouche and Oropouche-like viruses carrying the Oropouche S segment, as well as the Mayaro virus, using a multiplexed one-step reverse transcription real-time polymerase chain reaction (RT-qPCR). A chimeric plasmid containing both Mayaro and Oropouche targets was designed and evaluated for the in vitro production of transcribed RNA, which could be easily used as a non-infectious external control. To track false-negative results due to PCR inhibition or equipment malfunction, the MS2 bacteriophage was also included in the multiplex assay as an internal positive control. The specificity of the multiplex assay was evaluated by Primer-Blast analysis against the entire GenBank database, and further against a panel of 17 RNA arboviruses. The results indicated an accurate and highly sensitive assay with amplification efficiency greater than 98% for both targets, and a limit of detection between two and 20 copies per reaction. We believe that the assay described here will provide a tool for Mayaro and Oropouche virus detection, especially in areas where differential diagnosis of Dengue, Zika and Chikungunya viruses should be performed.

Mosquito-transmitted viruses - the great Brazilian challenge

ABSTRACT Arboviruses pose a serious threat to public health worldwide, overloading the healthcare system and causing economic losses. These viruses form a very diverse group, and in Brazil, arboviruses belonging to the families Flaviviridae and Togaviridae are predominant. Unfortunately, the number of arboviruses increases in proportion with factors such as deforestation, poor sanitation, climate changes, and introduction of new viruses like Chikungunya virus and Zika virus. In Brazil, dengue is endemic, along with the presence of other arboviruses. The situation is complicated by the scarcity of diagnostic infrastructure and the absence of approved vaccines for these diseases. Disease control, thus, relies solely on vector control. Therefore, enhanced clinical knowledge and improved general awareness about these arboviruses are indispensable to tackle diagnostic inadequacies.

Mayaro fever in an HIV-infected patient suspected of having Chikungunya fever

Abstract Arboviruses impose a serious threat to public health services. We report a case of a patient returning from a work trip to the Amazon basin with myalgia, arthralgia, fever, and headache. During this travel, the patient visited riverside communities. Both dengue and Chikungunya fevers were first suspected, tested for, and excluded. Mayaro fever was then confirmed by reverse transcription polymerase chain reaction followed by next-generation sequencing and phylogenetic reconstruction. The increased awareness of physicians and consequent detection of Mayaro virus in this case was only possible due a previous surveillance program with specific health personnel training about these neglected arboviruses.

Preparação e resposta à introdução do vírus Chikungunya no Brasil / Preparation and response to introduction of Chikungunya virus in Brazil / Preparación y respuesta a la introducción del virus Chikungunya en Brasil

O CHIKV é um vírus RNA que pertence ao gênero Alphavírus da família Togaviridae. O nome chikungunya deriva de uma palavra em Makonde que significa aproximadamente “aqueles que se dobram”, descrevendo a aparência encurvada de pacientes que sofrem de artralgia intensa. Casos humanos com febre, exantema e artrite aparentando ser CHIKV foram relatados no início de 1770. Porém, o vírus não foi isolado do soro humano ou de mosquitos até a epidemia na Tanzânia de 1952-53. Outros surtos ocorreram subsequentemente na África e na Ásia. Muitos ocorreram em pequenas comunidades ou comunidades rurais. No entanto,na Ásia, cepas de CHIKV foram isoladas durante grandes surtos urbanos em Bangkok eTailândia em 1960 e em Calcutá e Vellore, na Índia, durante as décadas de 60 e 70.

Chikungunya virus infection: report of the first case diagnosed in Rio de Janeiro, Brazil / Infecção pelo virus Chikungunya: relato do primeiro caso diagnosticado no Rio de Janeiro, Brasil

Initially diagnosed in Africa and Asia, the Chikungunya virus has been detected in the last three years in the Caribbean, Italy, France, and the United States of America. Herein, we report the first case for Rio de Janeiro, Brazil, in 2010.

Antes diagnosticado na África e na Ásia, o vírus Chikungunya foi detectado nos últimos três anos, no Caribe, na Itália, na França e nos Estados Unidos. Relatamos o primeiro caso do Rio de Janeiro, Brasil, em 2010.

Chikungunya virus outbreak in Kerala, India, 2007: a seroprevalence study

India was affected by a major outbreak of chikungunya fever caused by Chikungunya virus (CHIKV) during 2006-2007. Kerala was the worst affected state during 2007 with a contribution of 55.8 percent suspected cases in the country. However, except for clinically reported case records, no systematic information is available on infection status of CHIKV in the region. Hence, we carried out a post-epidemic survey to estimate seroprevalence status [immunoglobulin G (IgG)] in the community using commercially available indirect immunofluorescence test. This methodology had been reported to be highly specific and sensitive for CHIKV infection. The study area selected was the worst affected mid-highlands region of Kerala which harbour vast area of rubber plantations. The study evidenced 68 percent of the population to be seropositive for CHIKV IgG. Males were found more affected than females (χ2 = 9.86; p = 0.002). Among males, prevalence was significantly higher in the age classes 21-30 (χ2 = 5.46; p = 0.019) and 31-40 (χ2 = 5.84; p = 0.016) years. This may be due to high occupational risk of the male population engaged in plantation activities exposed to infective bites of Aedes albopictus. The current study provides an insight into the magnitude of CHIKV outbreak in Kerala.

Laboratory diagnosis of chikungunya virus: Do we really need it.

Chikungunya (CHIK) fever is a re-emerging Aedes mosquito-transmitted viral disease caused by CHIK virus belonging to the Togaviridae family of genus Alphavirus. The disease is almost self-limiting, occurs with characteristic triad of sudden onset fever, rash and arthritis. During the recent outbreak CHIKV was also found to cause long-term arthralgia, severe neurological disease and even fatalities. Although there are no antiviral or vaccines available for CHIKV, still there are several advantages to diagnose the infection. The present article provides an overview of various diagnostic modalities available and its significance by searching PubMed MeSH terms "Chikungunya virus" and "Diagnosis" for recent articles. The gold standard of CHIKV diagnosis is culture, yet requires facilities and skills. Highly sensitive and specific PCR assays for CHIKV have been developed, but the reagents and equipment are costly for widespread use. Serological diagnosis by detecting IgM antibody is most widely used as it is relatively cheaper and easier to perform. Disadvantages of antibody testing are cross-reactivity with other alpha viruses, cannot differentiate between recent past and acute infection, and its sensitivity varies in clinical settings. When tested for diagnosing acute CHIKV disease, sensitivities were just 4 to 22% and after 1 week rose to more than 80%. As most acutely infected patients seek medical attention within the first few days of illness, the ideal test should detect RNA or antigen. Therefore, the more realistic aim would be to develop a reliable antigen detection assay that could be used in rural areas, where CHIKV infection often occurs.

Rapid spread of Chikungunya virus infection in Orissa: India.

Background & objectives : A large number of cases of undiagnosed fever and joint pain were reported from different parts of the State of Orissa since February 2006. Epidemiological and laboratory investigation were carried out to confirm the cause of emerging illness, which was provisionally suspected as Chikungunya (CHIK) fever. Methods: Upon getting the reports of suspected CHIK like illness in different parts of the State, epidemic investigations were carried out in the outbreak affected villages. Case history was recorded, clinical examination undertaken and blood samples collected for seroconfirmation for CHIK IgM antibody using ELISA based kit. Simultaneously vector survey was also carried out. Results: With no previous record of CHIK infection in the State, the first outbreak was confirmed during February 2006. Subsequently, the infection spread to 13 of 30 districts in different episodes covering 79 villages till November 2007. Attack rate was 9-43 per cent in the different outbreaks with average seropositivity of 24 per cent to CHIK specific IgM. Morbidity was high though no deaths were recorded. Aedes aegypti and Ae. albopictus were identified as the possible vectors for transmission. Interpretation & conclusions : The report confirmed emergence of CHIK infection in the State of Orissa, India, and its spread to a larger geographic zone in a short period which warrants public health measures to control further spread.

Preparación y respuesta ante la eventual introducción del virus chikungunya en las Américas / Preparation and response to the eventual introduction of the virus Chikungunya in the Americas

Las siguientes guías fueron concebidas para ser adaptadas por cada País Miembro para mejorar los conocimientos sobre esta amenaza y para brindar las herramientas necesarias que permitan establecer las estrategias más adecuada para prevenir la importación de CHIKV a la Región, o para su control. Proporcionan orientación sobre cómo detectar un brote de la enfermedad, desarrollar las investigaciones epidemiológicas y prevenir o mitigar la diseminación de la enfermedad en la Región. Alentamos a las personas involucradas en la aplicación de estas guías a tener en cuenta todos los conocimientos disponibles y la capacidad propia de cada país para afrontar la eventual introducción del CHIKV. Se deben tomar medidas cuanto antes para poner en marcha las acciones necesarias para disminuir el impacto que este nuevo arbovirus que puede existir en nuestra Región.

Chikungunya virus iridocyclitis in Fuchs’ heterochromic iridocyclitis.

We are reporting a case of bilateral Fuchs’ heterochromic iridocyclitis with chikungunya virus infection in the left eye. A 20-year-old female was presented with a past history of fever suggestive of chikungunya with bilateral Fuchs’ heterochromic iridocyclitis and complicated cataract. She had a tripod dendritic pattern of keratic precipitates by confocal microscopy in the left eye with a stippled pattern of keratic precipitates in both eyes. The real-time polymerase chain reaction (RT-PCR) assay in the aqueous humor detected 98 copies/ml of chikungunya virus RNA. The patient underwent clear corneal phacoemulsification with in-the-bag intraocular lens implantation in the left eye with a good visual outcome. This is the first report where the presence of chikungunya virus RNA has been associated with a case of bilateral Fuchs’ heterochromic iridocyclitis.

A study on chikungunya outbreak during 2007 in Kerala, south India.

Background & objectives: The first chikungunya outbreak occurred in Kerala during 2006 affecting 14 districts, followed by another during May 2007 affecting almost whole of the State. Four of the worst affected districts viz, Pathanamthitta, Idukki, Kottayam and Thrissur were surveyed during 2007 to understand the magnitude of the problem of chikungunya fever, particularly clinical signs and symptoms. Methods: A total of 1265 persons from 310 houses were surveyed door-to-door in 20 different localities representing four affected districts. The history and examination findings from 354 clinically diagnosed chikungunya cases were recorded. The symptoms recorded were fever, headache, myalgia, arthralgia, itch/rash, oedema, eye congestion, eye pain, oral ulcers, distaste, nausea, vomiting and haemorrhage. Results: The major symptoms were fever (100%), headache (97.5%), arthralgia (99.4%) and myalgia (99.4%). A significant difference was observed in oedema, distaste, nausea and headache among different age groups and these symptoms were reported to be lower (12.2-89.8%) in younger age group than in older age group (90.4-100%). No genderwise difference was observed for any of the symptoms. In clinically diagnosed chikungunya cases higher age group (>35 years) found with higher rate of severity with symptoms of oedema, distaste, nausea and headache when compared with lower age group (1-35 yr). Interpretation & conclusions: Chikungunya invaded Kerala State for the first time in 2006 and continues to be a major vector borne disease in the State. The clinical symptoms in affected cases highlighted high fever, sever myalgia and prolonged arthralgia, with occasional history of skin itch/rash (petechiae).

A case of bilateral presumed chikungunya neuroretinitis.

Chikungunya fever is a relatively rare from of vector-borne viral fever caused by chikungunya virus and spread by bites of the Aedes aegypti and Aedes albopictus mosquito. Epidemics of chikungunya fever have been reported in the past from different parts of the world. Although the virus had been passive for quite some time, recent reports of outbreaks of chikungunya fever in several parts of Southern India have confirmed the re-emergence of this virus. Symptoms of this infection include abrupt onset of fever, chills, and headache, rash, severe joint pain, conjunctival injection and photophobia. Ocular manifestations have been recently reported with this infection. We report a case of a 48-year-old female patient, who presented with defective vision two weeks after a serology proven chikungunya infection. There was bilateral neuroretinitis with peripapillary cotton wool spots. These findings should be kept in mind as an ocular manifestation of chikungunya virus infection.

Clinical profile of Chikungunya in infants.

OBJECTIVE: To define the clinical manifestations of Chikungunya infection in infants. METHODS: The inclusion criteria was fever (defined as axillary temperature > 99.6 degrees F) with any one of the following features; seizure, loose stools, peripheral cyanosis, skin manifestations or pedal edema in children less than one year. Details of disease from onset of illness till admission were noted and a thorough clinical examination was done at the time of admission. Daily follow-up was performed and the serial order of appearance of clinical features was noted till complete recovery. The sera collected from patients after the 7th day of onset of fever was analyzed for specific chikungunya antibody by IgM antibody capture enzyme linked immunosorbent assay (ELISA). RESULTS: Fifty six (56) infants were laboratory confirmed for chikungunya, consisting of 34 (60.71%) males and 22 (39.29%) females. 4 (7.14%) infants were less than 1 month of age, 39 (69.64%) 2-6 months old and 13 (23.21%) 7-12 months old. Fever was invariably present, but associated constitutional symptoms in infants consisted of lethargy or irritability and excessive cry. The most characteristic feature of the infection in infants was acrocyanosis and symmetrical superficial vesicobullous lesions were noted in most infants. Erythematous asymmetrical macules and patches were observed which later progressed to morbiliform rashes. The face and oral cavity was spared in all observed patients. CONCLUSION: An entirely different spectrum of disease is seen in infants with chikungunya as compared to older children who need to be carefully observed for. The morbidity and mortality of the disease may be avoided by the rational use of drugs and close monitoring of all infants.

Bilateral retinitis following chikun- gunya fever.

A 35-year-old male with a history of chikungunya fever, presented with diminution of vision in the right eye of one-week duration. His best corrected visual acuity (BCVA) was counting fingers 2 meters and 20/20 (Snellens) in the right and left eyes respectively. A diagnosis of neuroretinitis was made in the right eye while left eye showed features of retinitis. ELISA (serum) and polymerase chain reaction (aqueous) were positive for herpes simplex virus. The lesions did not show any response to antiviral or steroid treatment and appeared to be self-limiting. At five months follow-up, lesions had resolved well with BCVA of 20/120 and 20/20 in the right and left eyes respectively.

Chikungunya infection in neonates.

We describe two neonates in whom chikungunya infection was confirmed by RNA PCR. Important clinical features include apnea, fever, erythematous maculo-papular rash and generalized hyperpigmentation.

Chikungunya fever: a re-emerging viral infection.

Chikungunya (CHIK) fever is a re-emerging viral disease characterized by abrupt onset of fever with severe arthralgia followed by constitutional symptoms and rash lasting for 1-7 days. The disease is almost self-limiting and rarely fatal. Chikungunya virus (CHIKV) is a RNA virus belonging to family Togaviridae, genus Alphavirus. Molecular characterization has demonstrated two distinct lineages of strains which cause epidemics in Africa and Asia. These geographical genotypes exhibit differences in the transmission cycles. In contrast to Africa where sylvatic cycle is maintained between monkeys and wild mosquitoes, in Asia the cycle continues between humans and the Aedes aegypti mosquito. CHIKV is known to cause epidemics after a period of quiescence. The first recorded epidemic occurred in Tanzania in 1952-1953. In Asia, CHIK activity was documented since its isolation in Bangkok, Thailand in 1958. Virus transmission continued till 1964. After hiatus, the virus activity re-appeared in the mid-1970s and declined by 1976. In India, well-documented outbreaks occurred in 1963 and 1964 in Kolkata and southern India, respectively. Thereafter, a small outbreak of CHIK was reported from Sholapur district, Maharashtra in 1973. CHIKV emerged in the islands of South West Indian Ocean viz. French island of La Reunion, Mayotee, Mauritius and Seychelles which are reporting the outbreak since February, 2005. After quiescence of about three decades, CHIKV re-emerged in India in the states of Andhra Pradesh, Karnataka, Maharashtra, Madhya Pradesh and Tamil Nadu since December, 2005. Cases have also been reported from Rajasthan, Gujarat and Kerala. The outbreak is still continuing. National Institute of Communicable Diseases has conducted epidemiological, entomological and laboratory investigations for confirmation of the outbreak. These have been discussed in detail along with the major challenges that the country faced during the current outbreak.

Expression and Evaluation of Chikungunya Virus E1 and E2 Envelope Proteins for Serodiagnosis of Chikungunya Virus Infection

PURPOSE: Chikungunya virus (CHIKV) causes endemic or epidemic outbreaks of CHIKV fever, which is a mosquitoe-transmitted viral disease in Africa, India, South-East Asia, and recently Southern Europe. Currently, serological diagnostic tests such as hemagglutination inhibition test (HI test), in-house IgM capture enzyme-linked immunosorbent assays (ELISA), and indirect immunofluorescence test were used for diagnosis of chikungunya fever, which are based on whole virus antigens. MATERIALS AND METHODS: CHIKV E1, and E2 envelope proteins for the CHIKV-specific serodiagnostic reagents for chikungunya fever were expressed in baculovirus expression system. The seroreactivity of recombinant CHIKV E1 and E2 envelope proteins were evaluated using sera panels of patients from Laboratoire Marcel Merieux by indirect IgM capture ELISA. RESULTS: The recombinant CHIKV E1 and E2 envelope protein showed sensitivity of 77.5% and 90%, respectively. The specificities of both CHIKV E1 and E2 envelope proteins were 100%. CONCLUSION: The recombinant CHIKV E1 and E2 envelope proteins could be a useful diagnostic reagent for CHIKV infection.