Mosquito Cell Cultures and the Study of Arthropod-Borne Togaviruses

Igarashi, Akira

doi:10.1016/s0065-3527(08)60447-9

Cited by 19 publications

(12 citation statements)

References 117 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…20 Viral isolation and identification. Tissue culture flasks (25 cm 3 ) containing Aedes albopictus C6/36 cells 21 were inoculated with serum from suspected patients (diluted 1/20 in sterile phosphate-buffered saline) and incubated at 34ЊC for 10 days in minimal essential medium (Gibco-BRL) containing Earle's salts, L-glutamate, non-essential amino acids, 0.11% sodium bicarbonate, 105 U/ml of penicillin, 75 U/ml of streptomycin, and 2% fetal bovine serum. The cells were fixed with acetone (stored at Ϫ20ЊC) on slides to perform indirect immunofluorescence using the technique described by Gubler and others.…”

Section: Methodsmentioning

confidence: 99%

Application of molecular typing techniques in the 1998 dengue epidemic in Nicaragua.

Balmaseda¹,

Sandoval²,

Pérez³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

Abstract. This report presents the results of applying the reverse transcriptase-polymerase chain reaction (RT-PCR) to the analysis of clinical specimens during the 1998 dengue epidemic in Nicaragua. The RT-PCR was validated through comparison with viral isolation, resulting in a sensitivity of 100% and a specificity of 90%. In-country application of the RT-PCR permitted the rapid identification of dengue-3 virus as the cause of the epidemic at the beginning of 1998 and the detection of the reintroduction of dengue-2 virus in the middle of the year. Nineteen isolates of dengue-3 and one of dengue-2 were characterized using the restriction site-specific (RSS)-PCR technique. This showed that the dengue-3 strain belonged to the ''Sri Lanka'' subtype and that the dengue-2 strain belonged to the ''Jamaica'' subtype, both of which have been associated with hemorrhagic dengue in the Americas. The application of these simple PCR-based strain typing methods in a country endemic for dengue virus infections can help to characterize the transmission dynamics of this important emerging infectious disease problem and provide this information to local health authorities in a timely manner so that appropriate control measures can be implemented.

show abstract

Section: Methodsmentioning

confidence: 99%

Application of molecular typing techniques in the 1998 dengue epidemic in Nicaragua.

Balmaseda¹,

Sandoval²,

Pérez³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

show abstract

“…[15][16][17] After 7 days, supernatant was separated from each culture vial at 1500 rpm for 10 minutes, 4° C and stored at -70° C, to be used as positive controls. The presence of dengue serotypes was confi rmed by multiplex RT-PCR, using four serotype specifi c primers pairs described earlier.…”

Section: Detection Of Dengue Serotypes By Multiplex Rt-pcrmentioning

confidence: 99%

Correlation of disease spectrum among four Dengue serotypes: a five years hospital based study from India

Kumaria¹

2010

Braz J Infect Dis

View full text Add to dashboard Cite

The recognition of DF (DHF Dengue Hemorrhagic Fever) is very complicated due to occurrence of a wide spectrum of clinical signs and symptoms during acute phase of illness. Moreover, presence of four serotypes further complicates the prognosis. To investigate the predictors of disease severity and elucidate the prognostic markers among four dengue serotypes, this study was conducted on 320 inpatients having acute febrile illness clinically suspected as DI, over a period of fi ve years. Dengue serotypes were confi rmed by multiplex reverse transcriptase (RT)-PCR. Eighty patients were positive for DI with presence of Den-1, Den-2, Den-3, and Den-4 in 8, 35, 27 and 10 patients, respectively. The severe clinical manifestations, abdominal pain and hepatomegaly, were comparatively higher in Den-2 patients. Liver aminotransferases levels were also higher in Den-2 patients (app. 5 fold). This study clearly indicates the hyperendemicity of all dengue serotypes. Nucleotide sequencing of Envelope region revealed that the presently emerged Den-3 belongs to type III, having high homology with genotype responsible for number of outbreaks in 1980s. The re-emergence of this deadly type can be suspected to cause more outbreaks in future and is a matter of great concern.

show abstract

“…Los cultivos celulares de insectos se utilizan, comúnmente, como sustratos para el aislamiento e identificación de arbovirus (18); sin embargo, existe en la actualidad una amplia gama de aplicaciones de esta técnica en investigaciones biomédicas y tecnológicas (19), entre las cuales sobresalen: la expresión de genes exógenos a partir de moléculas de ADN recombinante para el análisis de procesos genéticos de regulación, los cuales son difíciles de medir en el organismo entero (20); los estudios de cocultivos con parásitos (21); los estudios de entomopatógenos, reguladores del crecimiento o de compuestos bioquímicos usados contra insectos (22); la detección y dosis de entomopatógenos o sus toxinas de muestras ambientales (23); la respuesta hormonal in vitro (24); la fisiología celular (25), y el aislamiento y la caracterización de bioinsecticidas (26).…”

unclassified

Características de nuevos cultivos celulares derivados de tejidos embrionarios de Aedes aegypti (Diptera: Culicidae).

Silva¹,

Escovar²,

Bello³

2005

biomedica

View full text Add to dashboard Cite

Introducción. Los cultivos celulares de insectos son una metodología útil en estudios biomédicos y tecnológicos. Objetivo. El propósito principal del presente trabajo fue obtener y caracterizar cultivos celulares derivados de tejidos embrionarios de Aedes aegypti. Materiales y métodos. Se emplearon huevos embrionados para los explantes de tejidos en los medios de cultivos MM/VP12 y L-15/Grace, con suplemento de 20% de suero fetal bovino y una mezcla al 1% de antibiótico y antimicótico, con un rango de pH entre 6,8 y 7,0. Los cultivos se incubaron a una temperatura de 28 o C sin atmósfera de CO 2 . Resultados. El crecimiento celular se obtuvo en el medio L-15/Grace, 3 semanas después de haber sido sembrados los tejidos embrionarios; sin embargo, se necesitaron 6 meses para la formación de la monocapa confluente. Desde agosto de 2003 hasta junio de 2004, se habían realizado 28 subcultivos. Las células se caracterizaron morfológicamente; predominaron las formas epitelioides en subcultivos de pases altos. También se reconocieron las particularidades morfométricas del cariotipo y, además, se determinaron los perfiles isoenzimáticos y moleculares de los cultivos celulares, los cuales se compararon con muestras de adultos de la especie tomadas de la misma colonia y con líneas celulares derivadas de otros insectos. Discusión. Estas células representan, potencialmente, un importante sistema in vitro en investigaciones básicas y aplicadas. Palabras clave: Aedes aegypti, cultivos celulares, vesículas, cariotipos, isoenzimas, RAPD-PCR.Characteristics of new cell cultures derived from embryonic tissues of Aedes aegypti (Diptera: Culicidae) Introduction. Cell cultures from insects are a useful methodology in technological and biomedical studies.Objective. The present work was aimed at obtaining and characterizing cell cultures derived from Aedes aegypti embryonic tissues. Materials and methods. Embryonated eggs were used for embryonic tissue explants in L-15/ Grace and MM/VP12 culture media, supplemented with 20% fetal bovine serum and a mixture of 1% antimycotic and antibiotics, at a pH ranging from 6.8 to 7.0. The incubation temperature was 28 o C; a CO 2 atmosphere was not required. Results. Cell growth was obtained in L-15/Grace medium three weeks after embryonic tissues explants. Six months were required for achieving a confluent monolayer. Twenty-eight serial cell subcultures were carried out from August 2003 to June 2004. Cell morphology was characterized as epithelial in the later subcultures. Karyotype morphometry as well as molecular and isozymatic profiles were established. The cultures were compared with adult samples from the species taken from the same colony and with cell lines derived from other insects.

show abstract

Mosquito Cell Cultures and the Study of Arthropod-Borne Togaviruses

Cited by 19 publications

References 117 publications

Application of molecular typing techniques in the 1998 dengue epidemic in Nicaragua.

Application of molecular typing techniques in the 1998 dengue epidemic in Nicaragua.

Correlation of disease spectrum among four Dengue serotypes: a five years hospital based study from India

Características de nuevos cultivos celulares derivados de tejidos embrionarios de Aedes aegypti (Diptera: Culicidae).

Contact Info

Product

Resources

About