RESUMO
Enzyme-linked immunosorbent assays have been widely used for diagnosis of FeLV and feline immunodeficiency virus (FIV) infections. Various ELISA kits for FeLV are available from several manufacturers. Although these tests are configured in a variety of formats, they are all direct antigen-detection systems for the viral core protein p27. On the other hand, ELISA for FIV exposure detects specific feline antibody to FIV. Basic immunoassay principles and the application of ELISA technology used in FeLV and FIV ELISA kits are described.
Assuntos
Ensaio de Imunoadsorção Enzimática , Vírus da Imunodeficiência Felina/isolamento & purificação , Infecções por Lentivirus/veterinária , Vírus da Leucemia Felina/isolamento & purificação , Leucemia Felina/diagnóstico , Animais , Anticorpos Antivirais/sangue , Antígenos Virais/sangue , Gatos , Vírus da Imunodeficiência Felina/imunologia , Infecções por Lentivirus/diagnóstico , Vírus da Leucemia Felina/imunologia , Valor Preditivo dos Testes , Kit de Reagentes para Diagnóstico/veterináriaAssuntos
Vírus da Imunodeficiência Felina/imunologia , Infecções por Lentivirus/veterinária , Vírus da Leucemia Felina/imunologia , Leucemia Felina/epidemiologia , Fatores Etários , Animais , Anticorpos Antivirais/sangue , Antígenos Virais/sangue , Castração/veterinária , Gatos , Ensaio de Imunoadsorção Enzimática , Feminino , Infecções por Lentivirus/complicações , Infecções por Lentivirus/epidemiologia , Leucemia Felina/complicações , Masculino , Prevalência , Fatores de Risco , Fatores Sexuais , Estados Unidos/epidemiologiaRESUMO
The feline immunodeficiency virus (FIV) is a recently identified feline lentivirus that has been found at significant levels in domestic cat populations worldwide. A microdilution plate format, monoclonal antibody-based enzyme-linked immunosorbent assay was developed for the detection of the FIV group-associated antigen (gag) designated p24. Assays of serially diluted samples containing disrupted virus showed that the assay had a sensitivity limit of approximately 0.2 ng/ml for FIV p24. The assay was approximately eightfold more sensitive than the assay for viral reverse transcriptase activity when it was tested with diluted tissue culture samples. A qualitative confirmation assay by standard antibody inhibition techniques was coupled to the screening test methodology. The test was used to detect and confirm the presence of virus in cultured feline lymphocytes from infected animals.
Assuntos
Antígenos Virais/análise , Ensaio de Imunoadsorção Enzimática , Retroviridae/imunologia , Proteínas do Core Viral/imunologia , Animais , Anticorpos Monoclonais , Anticorpos Antivirais , Gatos , Estudos de Avaliação como Assunto , Infecções por Retroviridae/diagnósticoRESUMO
The feline T-cell lymphotropic lentivirus (feline immunodeficiency virus) is a recently described feline-specific retrovirus that can produce chronic immunodeficiency-like disorders in cats. A microdilution plate format enzyme-linked immunosorbent assay has been developed to detect the presence of antibody to the virus in feline serum or plasma. Temporal studies performed with experimentally infected animals show that seroconversion can be demonstrated 3 to 4 weeks after exposure to the virus. Results of a serosurvey (n = 1,556 samples) indicate that infection is fairly common in both clinic (5.2%) and sick cat (15.2%) populations. Western blot (immunoblot) and sodium dodecyl sulfate radioimmunoprecipitation assays were developed to confirm microdilution plate test results and to identify peptides specific for the feline immunodeficiency virus. All microdilution plate test positive results and selected negative results were confirmed by one or both of these procedures. These data demonstrate that this microassay plate enzyme-linked immunosorbent assay is a very sensitive and specific test for detection of antibody to the feline immunodeficiency virus.
Assuntos
Anticorpos Antivirais/análise , Doenças do Gato/diagnóstico , Ensaio de Imunoadsorção Enzimática , Infecções por Retroviridae/veterinária , Retroviridae/imunologia , Animais , Western Blotting , Gatos , Eletroforese em Gel de Poliacrilamida , Testes de Precipitina , Valor Preditivo dos Testes , Infecções por Retroviridae/diagnósticoRESUMO
Treatment of mouse mammary glands with a high concentration of 7,12-dimethylbenzo(a)anthracene in whole organ culture was reported by Banerjee et al. to transform foci of lobuloalveoli to a hormone-independent state, and to give rise to mammary hyperplastic outgrowths and adenocarcinomas in vivo. In the present study using the identical system, mammary glands of BALB/c mice were exposed to 7,12-dimethylbenzo(a)anthracene or N-2-fluorenylacetamide at low concentrations that bring about maximal incidences of the hormone-independent hyperplastic lobuloalveolar lesions with minimal cytotoxicity. After morphological development of the lobuloalveoli in culture, the glands were enzymatically dissociated into cells and inoculated into gland-free inguinal mammary fat pads of syngeneic mice bearing pituitary gland implants during the initial 8 weeks. After 11 months, fragments of the resultant mammary outgrowths from each mouse were implanted into the gland-free inguinal mammary fat pads of 3 syngeneic mice (not bearing pituitary gland supplements) and were permitted to grow for another 11 months. Mammary outgrowths from the primary and secondary implants were neither neoplastic, anaplastic, nor dysplastic. Also, no hyperplasia in any mammary outgrowth could be attributed to the action of either carcinogen, especially when outgrowths were compared with contralateral outgrowths that arose from the control glands exposed to dimethyl sulfoxide (solvent of the carcinogens) in culture and/or with untreated thoracic mammary glands of the same hosts. One interpretation of these findings is that the hormone-independent, hyperplastic alveolar lesions may not be an appropriate in vitro marker of oncogenic transformation by chemical carcinogens in culture. The great variety of procarcinogens and activated carcinogens that bring about this lesion in vitro and its morphological similarity to presumptive mammary preneoplastic lesions in vivo weigh against this interpretation. A second hypothesis is that high concentrations of procarcinogens, despite their considerable cytotoxicity, complete a multistep process of oncogenic transformation in surviving mammary epithelium, whereas low concentrations optimized to produce the lesions in maximal number do not.
Assuntos
2-Acetilaminofluoreno/toxicidade , 9,10-Dimetil-1,2-benzantraceno/toxicidade , Benzo(a)Antracenos/toxicidade , Glândulas Mamárias Animais/efeitos dos fármacos , Neoplasias Mamárias Experimentais/induzido quimicamente , Animais , Relação Dose-Resposta a Droga , Hiperplasia/induzido quimicamente , Glândulas Mamárias Animais/patologia , Glândulas Mamárias Animais/transplante , Camundongos , Camundongos Endogâmicos BALB C , Técnicas de Cultura de ÓrgãosAssuntos
Carcinógenos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Glândulas Mamárias Animais/efeitos dos fármacos , Neoplasias Mamárias Experimentais/induzido quimicamente , Vírus do Tumor Mamário do Camundongo/efeitos dos fármacos , Ativação Viral/efeitos dos fármacos , 2-Acetilaminofluoreno/farmacologia , 9,10-Dimetil-1,2-benzantraceno/farmacologia , Animais , Transformação Celular Viral , Células Cultivadas , DNA Viral/biossíntese , Feminino , Neoplasias Mamárias Experimentais/microbiologia , Vírus do Tumor Mamário do Camundongo/genética , Camundongos , Camundongos Endogâmicos , RNA Viral/análise , Proteínas Virais/análiseAssuntos
Bucladesina/farmacologia , Diferenciação Celular/efeitos dos fármacos , Fator de Crescimento Epidérmico/farmacologia , Glândulas Mamárias Animais/fisiologia , Prostaglandinas/farmacologia , Animais , Caseínas/biossíntese , Células Cultivadas , AMP Cíclico/metabolismo , Feminino , Glândulas Mamárias Animais/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Peso MolecularRESUMO
The mouse mammary gland in serum-free whole organ culture can be manipulated hormonally to undergo one complete physiological cycle consisting of lobuloalveolar development, functional differentiation and regression, mimicking processes that occur in vivo. A second cycle has not previously been achieved in vitro. The present study has identified a specific requirement for epidermal growth factor (EGF) in the morphological development of mammary lobuloalveoli, allowing two complete cycles of development and regression in culture.