Fluorescent Antibody Test for Rapid Detection of West Nile Virus Antigen in Avian Tissues

West Nile virus was recovered from the brain of a red-tailed hawk that died in Westchester County, N.Y., in February 2000. Multiple foci of glial cells, lymphocytes, and a few pyknotic nuclei were observed in the brain. Three to 4 days after inoculation of Vero cells with brain homogenates, cytopathic changes were detected. The presence of West Nile virus antigen in fixed cells or cell lysates was revealed by fluorescent antibody testing or enzyme-linked immunosorbent assay, respectively. Furthermore, Reverse transcriptase-PCR with primers specific for the NS3 gene of West Nile virus resulted in an amplicon of the expected size (470 bp). Electron microscopy of thin sections of infected Vero cells revealed the presence of viral particles approximately 40 nm in diameter, within cytoplasmic vesicles. The demonstration of infection with the West Nile virus in the dead of the winter, long after mosquitoes ceased to be active, is significant in that it testifies to the survival of the virus in the region beyond mosquito season and suggests another route of transmission: in this case, prey to predator.

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An indirect fluorescent antibody test for the rapid detection of infectious pancreatic necrosis virus in tissues

Journal of Fish Diseases ◽

10.1111/j.1365-2761.1981.tb01138.x ◽

1981 ◽

Vol 4 (4) ◽

pp. 309-315 ◽

Cited By ~ 13

Author(s):

R. N. SWANSON ◽

J. H. GILLESPIE

Keyword(s):

Rapid Detection ◽

Pancreatic Necrosis ◽

Indirect Fluorescent Antibody Test ◽

Fluorescent Antibody ◽

Antibody Test ◽

Infectious Pancreatic Necrosis Virus ◽

Indirect Fluorescent Antibody ◽

Necrosis Virus ◽

Infectious Pancreatic Necrosis ◽

Pancreatic Necrosis Virus

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Analysis of the Results of Monitoring Arbovirus Infections in the Volgograd Region in 2019

Epidemiology and Vaccinal Prevention ◽

10.31631/2073-3046-2021-20-1-51-59 ◽

2021 ◽

Vol 20 (1) ◽

pp. 51-59

Author(s):

A. O. Negodenko ◽

E. V. Molchanova ◽

D. R. Prilepskaya ◽

P. Sh. Konovalov ◽

O. A. Pavlyukova ◽

...

Keyword(s):

West Nile Virus ◽

Sindbis Virus ◽

Hemorrhagic Fever ◽

Virus Antigen ◽

Farm Animals ◽

West Nile ◽

Hemorrhagic Fever Virus ◽

Blood Sucking ◽

Volgograd Region ◽

California Serogroup

Relevance. Natural and climatic conditions, a variety of species composition of arthropods and vertebrates determine the possibility of circulation of arboviruses in the Volgograd region. The existence of natural foci of some arbovirus infections and the possibility of the formation of others suggests the need for annual monitoring of the causative agents of arbovirus diseases. Аim. Evaluation of the results of monitoring of arbovirus infections in the Volgograd region in 2019.Materials and methods: 806 blood serum samples from donors, 44 blood serum samples from febrile sick people, 300 blood serum samples from horses and 94 pools of blood-sucking mosquitoes were examined by immunofernal analysis. Result of the study of serum samples from donors in the Volgograd region, in 140 (17.4%) of 806 were found to have antibodies to the pathogen of West Nile fever (in 35 (4.3%) – IgM, in 105 (13.0%) – IgG), in 7 (2.2%) of 319 – to the Crimean hemorrhagic fever virus (in 4 (1.3%) – IgM, in 3 (0.9%) –IgG), and in 7 (2.9%) of 240 – IgG to the viruses of the California serogroup. Specific antibodies against viruses of Sindbis, Batai and Uukuniemi in the samples was not detected. The largest number of positive samples with the presence of IgG and IgM to the West Nile virus was found among residents of Volgograd (61 out of 240, 25.4%) and Volzhsky (25 out of 100, 25, 0%). Among 44 blood serums of febrile patients, 1 sample (2.3%) was found to contain an antigen of the Sindbis virus, and 2 samples (4.5%) – antigens California serogroup viruses. Specific immunoglobulins against West Nile virus were detected in 84 (28%) of 300 blood serums of farm animals (horses). In the study of 94 samples of field material (blood-sucking mosquitoes), West Nile virus antigen was detected in 14 (14.9%), Sindbis virus – in one sample (1.0%), Batai virus – in four samples (4.2%). Conclusions: the obtained results, along with the circulation of West Nile virus and Crimean hemorrhagic fever virus virus in the Volgograd region, indicate the presence of Sindbis, Batai and California serogroup viruses and necessitate further study of their role in the infectious pathology of the population.

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Detection of Rabies Virus Antigen in Dog Saliva Using a Latex Agglutination Test

Journal of Clinical Microbiology ◽

10.1128/jcm.38.8.3098-3099.2000 ◽

2000 ◽

Vol 38 (8) ◽

pp. 3098-3099 ◽

Cited By ~ 25

Author(s):

S. Kasempimolporn ◽

W. Saengseesom ◽

B. Lumlertdacha ◽

V. Sitprija

Keyword(s):

Rabies Virus ◽

Gamma Globulin ◽

Fluorescent Antibody ◽

Antibody Test ◽

Virus Antigen ◽

Latex Agglutination ◽

Agglutination Test ◽

Glass Slide ◽

Human Rabies ◽

Latex Particles

Dog bites are responsible for more than 90% of human rabies deaths in Asia. We developed a simple and inexpensive test based on latex agglutination (LA) for rabies virus antigen detection in dog saliva. Rabies virus antigen could be detected by agglutination on a glass slide using latex particles coated with gamma globulin. By evaluation of paired saliva-brain specimens from 238 dogs, the LA test using saliva was 99% specific and 95% sensitive compared to the fluorescent antibody test (FAT) on brain smears. The advantages of the LA test over the standard FAT are that it is comparatively simple and there is no need to kill the animal before examination.

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Comparison of three enzyme-linked immunosorbent assays and a direct fluorescent-antibody test for detection of respiratory syncytial virus antigen.

Journal of Clinical Microbiology ◽

10.1128/jcm.26.2.377-379.1988 ◽

1988 ◽

Vol 26 (2) ◽

pp. 377-379 ◽

Cited By ~ 4

Author(s):

L R Krilov ◽

L Marcoux ◽

H D Isenberg

Keyword(s):

Respiratory Syncytial Virus ◽

Fluorescent Antibody ◽

Antibody Test ◽

Virus Antigen ◽

Direct Fluorescent Antibody ◽

Respiratory Syncytial Virus Antigen ◽

Immunosorbent Assays ◽

Syncytial Virus

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Pathogenesis of Two Western Mediterranean West Nile Virus Lineage 1 Isolates in Experimentally Infected Red-Legged Partridges (Alectoris rufa)

Pathogens ◽

10.3390/pathogens10060748 ◽

2021 ◽

Vol 10 (6) ◽

pp. 748

Author(s):

Virginia Gamino ◽

Elisa Pérez-Ramírez ◽

Ana Valeria Gutiérrez-Guzmán ◽

Elena Sotelo ◽

Francisco Llorente ◽

...

Keyword(s):

Viral Load ◽

West Nile Virus ◽

Virus Antigen ◽

Western Mediterranean ◽

Post Inoculation ◽

Alectoris Rufa ◽

West Nile ◽

Neuronal Necrosis ◽

Molecular Determinants ◽

Bird Mortality

West Nile virus (WNV) is the most widespread flavivirus in the world with a wide vertebrate host range. Its geographic expansion and activity continue to increase with important human and equine outbreaks and local bird mortality. In a previous experiment, we demonstrated the susceptibility of 7-week-old red-legged partridges (Alectoris rufa) to Mediterranean WNV isolates Morocco/2003 and Spain/2007, which varied in virulence for this gallinaceous species. Here we study the pathogenesis of the infection with these two strains to explain the different course of infection and mortality. Day six post-inoculation was critical in the course of infection, with the highest viral load in tissues, the most widespread virus antigen, and more severe lesions. The most affected organs were the heart, liver, and spleen. Comparing infections with Morocco/2003 and Spain/2007, differences were observed in the viral load, virus antigen distribution, and lesion nature and severity. A more acute and marked inflammatory reaction (characterized by participation of microglia and CD3+ T cells) as well as neuronal necrosis in the brain were observed in partridges infected with Morocco/2003 as compared to those infected with Spain/2007. This suggests a higher neurovirulence of Morocco/2003, probably related to one or more specific molecular determinants of virulence different from Spain/2007.

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Use of an Immunoperoxidase Test for the Detection of Bovine Herpesvirus-1 in Aborted Fetal Tissue

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063878900100112 ◽

1989 ◽

Vol 1 (1) ◽

pp. 39-44 ◽

Cited By ~ 19

Author(s):

George H. Smith ◽

James K. Collins ◽

Jane Carman

Keyword(s):

Fluorescent Antibody ◽

Bovine Herpesvirus ◽

Antibody Test ◽

Fetal Tissue ◽

Virus Antigen ◽

Bovine Herpesvirus 1 ◽

Induced Abortions ◽

Formalin Fixed Paraffin Embedded ◽

Herpesvirus 1 ◽

Indirect Immunoperoxidase

An indirect immunoperoxidase (IP) procedure using a specific monoclonal antibody and an avidinbiotin-peroxidase complex was developed and applied to detect virus antigen in formalin-fixed, paraffin-embedded tissue sections. This IP procedure was compared with currently used diagnostic tests for detection of virus-induced abortions caused by bovine herpesvirus-1 (BHV–1). The IP procedure was applied to detect BHV-1 antigen in sections of liver and lung from 87 aborted fetuses. Sixteen of these cases were positive for viral antigen by IP staining. Sections from both liver and lung were positive in 15 of the 16 cases. A fluorescent antibody test (FA), which was applied to acetone-fixed frozen sections of liver and lung, gave positive results on 12 of the 87 fetuses, 11 of which were also positive by IP. Seven of the 12 FA-positive cases were positive on both sections of liver and lung. When FA and IP were compared, FA had a sensitivity of 67% and IP had a sensitivity of 94%. Virus was isolated from one of the 67 cases tested. The tissues in which antigen was most frequently detected by IP were liver, lung, and kidney. Distinct multifocal staining was seen in positive sections of all these tissues.

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