Detection of Bovine Respiratory Syncytial Virus using a Heterologous Antigen-Capture Enzyme Immunoassay

Based on the marked antigenic similarities that exist between antigens of the human and bovine strains of respiratory syncytial virus (RSV), an enzyme immunoassay (EIA) designed to detect human RSV was used to detect bovine RSV. The commercial test kit (RSV EIA) consists of a solid phase (beads) coated with a capture antiserum prepared against the Long strain of human RSV. The RSV EIA test was compared with the method of inoculation of cell cultures and fluorescent antibody (FA) staining of lung tissue for the detection of bovine RSV. Using a cell culture-propagated stock of strain 375 of bovine RSV, the threshold of sensitivity of the EIA test for the cattle strain of RSV was determined to be ≤ 102,3 CCID50/ml. In addition, RSV EIA detected the bovine RSV in nasal samples obtained from 3 experimentally inoculated cattle. The RSV EIA exhibited a sensitivity of ≥ 80% during the period that shedding of infectious virus took place. All of the bovine RSV FA-positive lung samples (n = 37) were positive by the RSV EIA. Twenty-six of the remaining 214 bovine RSV FA-negative lung samples were positive by the RSV EIA. The RSV EIA was also used to test 137 nasal swabs obtained from cases of bovine respiratory disease. Of these, 38 tested positive by RSV EIA. All samples that tested positive by EIA were confirmed by blocking assays using hyperimmune serum anti-bovine RSV and a pool of monoclonal antibodies specific for that virus.

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2620. Respiratory Syncytial Virus Rapid Antigen Detection Test, Can It Be Trusted?

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.2298 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S912-S912

Author(s):

Nicole Titze ◽

Jasjit Singh ◽

Wendi Gornick

Keyword(s):

Respiratory Syncytial Virus ◽

Clinical Decision Making ◽

Fluorescent Antibody ◽

Antigen Detection ◽

Urgent Care ◽

Viral Fusion ◽

Rapid Antigen Detection Test ◽

Viral Fusion Protein ◽

Test Kit ◽

Syncytial Virus

Abstract Background Many emergency departments and urgent care settings use the commonly available Respiratory Syncytial Virus Rapid Antigen Detection Test (RSV RADT) to diagnose children with RSV. We noted discordant results between RADT and definitive testing. Our study looked at the positive predictive value (PPV) and the false discovery rate (FDR) of the RSV RADT at our facility. Methods We pro- and retrospectively reviewed all patients with positive RSV RAPD tests from July 1, 2017 through March 31, 2019. The test utilized was the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA), which detects the viral fusion protein present in RSV. Of the tests performed, we chose patients who had definitive testing with either a direct fluorescent antibody (DFA) or a polymerase chain reaction (PCR). We then calculated the PPV as well as the FDR of the RSV RADT during the total interval period, as well as off-season periods (April 1 through October 31) and in-season periods (November 1 through March 31). Results During the study period there were 1128 RSV RADT tests performed, of which 232 had definitive testing with either DFA or PCR (Figures 1 and 2). We found the overall PPV during the study period was 63.3%. During the off-season 30 positive RSV RADT received definitive testing, of which 6 were positive, which yields a PPV of only 20%. In season, 202 RSV RADT received additional testing with 141 positive for RSV. The PPV was 69.8%. The FDR correlated with 36.7% throughout the entire studied period, 80% during the off-season and 30.2% during in-season. As expected, the PPV was higher during times of higher prevalence (Figure 3). Conclusion Based on our results, utilization of the RSV RADT during time of low prevalence yields a high false detection rate and should therefore be discouraged. The use during times of high prevalence yields only modest results and is unlikely to aid in clinical decision-making. Our results differ from those published by the manufacturer (PPV 84%), and may reflect differences in sample collection in the acute care setting. Disclosures All authors: No reported disclosures.

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The use of cough/nasal swabs in the rapid diagnosis of respiratory syncytial virus infection by the fluorescent antibody technique

Journal of Hygiene ◽

10.1017/s0022172400028746 ◽

1970 ◽

Vol 68 (2) ◽

pp. 283-292 ◽

Cited By ~ 5

Author(s):

Joyce McQuillin ◽

P. S. Gardner ◽

Patricia M. sturdy

Keyword(s):

Respiratory Syncytial Virus ◽

Virus Infection ◽

Nasal Swab ◽

Fluorescent Antibody ◽

Rapid Diagnosis ◽

Fluorescent Antibody Technique ◽

Antibody Technique ◽

Nasal Swabs ◽

Respiratory Virus Infection ◽

Syncytial Virus

SUMMARYThirty-five consecutive infants admitted into hospital in Newcastle upon Tyne with acute respiratory disease had cough/nasal swabs and nasopharyngeal secretions taken. Both types of specimens were examined the fluorescent antibody technique for respiratory syncytial virus; isolation techniques were also used. Twenty-eight specimens of nasopharyngeal secretion were positive, as were 26 of the corresponding cough/nasal swab preparations. Respiratory syncytial virus was isolated from all but one.Sixteen consecutive children who were only suitable for examination cough/nasal swab preparations were also investigated isolation and fluorescent antibody techniques for respiratory syncytial virus. Respiratory syncytial virus was isolated from eight, seven of whom were positive the fluorescent antibody technique. The use of cough/nasal swab preparations stained the fluorescent antibody technique, although not as efficient as nasopharyngeal secretions, may have a place in the rapid diagnosis of respiratory virus infection in older children and children in general practice. The importance of rapid diagnosis for respiratory virus infection in relationship to antiviral therapy was also discussed.

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A comparative study of methods for the diagnosis of respiratory virus infections in childhood

Epidemiology and Infection ◽

10.1017/s002217240004211x ◽

1969 ◽

Vol 67 (4) ◽

pp. 659-670 ◽

Cited By ~ 26

Author(s):

Patricia M. Sturdy ◽

Joyce McQuillin ◽

P. S. Gardner

Keyword(s):

Tissue Culture ◽

Respiratory Syncytial Virus ◽

Virus Infection ◽

Respiratory Syncytial Virus Infection ◽

Fluorescent Antibody ◽

Health Department ◽

Fluorescent Antibody Technique ◽

Antibody Technique ◽

Nasal Swabs ◽

Syncytial Virus

SUMMARYNasopharyngeal secretions and cough/nasal swabs were taken from 111 children admitted to hospital in Newcastle upon Tyne with acute lower respiratory disease. A comparison was made between nasopharyngeal secretions and cough/nasal swabs as material for isolation of viruses in tissue culture. These results were, in turn, compared with those obtained by applying a fluorescent antibody technique to the exfoliated cells in the nasopharyngeal secretions for the rapid diagnosis of respiratory syncytial virus infection.More viruses were isolated in tissue culture from nasopharyngeal secretion than from cough/nasal swabs. Further evidence for the superiority of nasopharyngeal secretions was obtained by comparing the virus isolations in the laboratory in 1967 with those in 1968. Respiratory syncytial virus was not only isolated more often but more quickly in tissue culture inoculated with nasopharyngeal secretions.The fluorescent antibody technique not only provided a diagnosis on the patient's day of admission in 95% of those infected with respiratory syncytial virus but also proved to be as sensitive as the culture of nasopharyngeal secretions and considerably more sensitive than the culture of cough/nasal swabs for the diagnosis of respiratory syncytial virus infection.We wish to acknowledge the invaluable help given us by consultants, medical officers, ward sisters and their staff in the paediatric wards of the Child Health Department. We also wish to thank the technical staff in the Department of Virology for their aid.

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A study of possible biohazards in the fluorescent antibody test using adenovirus, coxsackievirus, herpesvirus, and respiratory syncytial virus as antigens

Journal of Clinical Microbiology ◽

10.1128/jcm.4.4.322-325.1976 ◽

1976 ◽

Vol 4 (4) ◽

pp. 322-325

Author(s):

D Bardell

Keyword(s):

Respiratory Syncytial Virus ◽

Infectious Virus ◽

Fluorescent Antibody ◽

Antibody Test ◽

Adenovirus Type ◽

Antigenic Determinants ◽

Syncytial Virus ◽

Simplex Virus ◽

Adenovirus Type 5

Infectious adenovirus type 5 and coxsackievirus type B5, both nonlipid-containing viruses, were isolated from cells fixed in acetone at 22 degrees C for 15 min, from acetone used for fixation, from the solution used for washing slides during the fluorescent antibody procedure, and after complete processing of antigen preparations with serial twofold dilutions of human antisera and fluorescein-labeled goat anti-human immunoglobulin G. Lipid-containing herpes simplex virus type 1 and respiratory syncytial virus were inactivated by acetone, and infectious virus could not be recovered at any stage in the fluorescent antibody test. Fixation in acetone at 56 degrees C destroyed the infectivity of adenovirus 5 and coxsackievirus B5 within 30 min, but no adverse effect on the antigenic determinants of either virus occurred until after 60 min, thus demonstrating that these antigens can be utilized without the hazard of infectious virus.

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RS virus diagnosis: comparison of isolation, immunofluorescence and enzyme immunoassay

Memórias do Instituto Oswaldo Cruz ◽

10.1590/s0074-02761986000200013 ◽

1986 ◽

Vol 81 (2) ◽

pp. 225-232 ◽

Cited By ~ 8

Author(s):

Marilda M. Siqueira ◽

Vanja Ferreira ◽

Jussara P. Nascimento

Keyword(s):

Tissue Culture ◽

Respiratory Syncytial Virus ◽

Enzyme Immunoassay ◽

Bacterial Contamination ◽

Virus Isolation ◽

Rapid Diagnosis ◽

Tissue Cultures ◽

Virus Diagnosis ◽

Under Five ◽

Syncytial Virus

Two techniques for rapid diagnosis, immunofluorescence (IFAT) and enzyme immunoassay (EIA), have been compared with virus isolaion in tissue culture for the detection of respiratory syncytial virus (RSV) in specimens of nasopharyngeal secretions. The specimens were obtained from children under five years of age suffering from acute respiratory iliness, during a period of six months from January to June 1982. Of 471 specimens examined 54 (11.5%) were positive by virus isolation and 180 (38.2%) were positive by immunofluorescence. The bacterial contamination of inoculated tissue cultures unfortunately prevented the isolation of virus from many samples. Specimens from 216 children were tested to compare enzyme immunoassay and immunofluorescence. Of these 60 (27%) were positive by EIA and 121 (56%) were positive by IFAT. Our results suggest that the EIA technique although highly specific is rather insensitive. This may be because by the time these tests were done the originl nasopharyngeal secretions were considerably diluted and contained more mucus fragments than the call suspension used for IFAT. Of the three techniques, IFAT gives the best results although EIA may be useful where IFAT is not possible.

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