Use of Fluorescent-Antibody Staining of Cytocentrifuge-Prepared Smears in Combination with Cell Culture for Direct Detection of Respiratory Viruses

Doing, Kirk M.; Jerkofsky, Mary Ann; Dow, Elaine G.; Jellison, Jo Ann

doi:10.1128/jcm.36.7.2112-2114.1998

Cited by 31 publications

(10 citation statements)

References 13 publications

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“…Rapid diagnosis allows the prompt initiation of effective antiviral therapy, especially in immunocompromised patients. Viral culture, usually in combination with immunofluorescence, has been regarded as the gold standard for the laboratory diagnosis of respiratory viruses (Doing et al, 1998;Johnston and Siegel, 1991). However, these tests are not rapid and their clinical value is often limited.…”

Section: Introductionmentioning

confidence: 99%

The detection of influenza A and B viruses in clinical specimens using a quartz crystal microbalance

Hewa¹,

Tannock

Mainwaring³

et al. 2009

Journal of Virological Methods

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Current methods for the accurate diagnosis of influenza based on culture of the virus or PCR are highly sensitive and specific but require specialised laboratory facilities and highly trained personnel and, in the case of viral culture, can take up to 14 days to obtain a definitive result. In this study, a quartz crystal microbalance-based immunosensor (QCM) has been developed and its potential evaluated for the rapid and sensitive detection of both influenza A and B viruses in laboratory-cultured preparations and clinical samples. The effective limit for detection by QCM for stock preparations of both A/PR/8/34 and B/Lee/40 viruses was 1 x 10(4) pfu/mL, associated with observed frequency shifts of 30 (+/-5) and 37 (+/-6.5) Hz, respectively. Conjugation of 13 nm gold nanoparticles to the detecting antibody improved the mass sensitivity of the immunosensor, resulting in a 10-fold increase in sensitivity and a detection limit of 1 x 10(3) pfu/mL for both preparations, with resulting frequency shifts of 102 (+/-11) and 115 (+/-5) Hz, respectively. Detection of virus in nasal washes with this technique was achieved by overnight passage in MDCK cultures prior to analysis. A comparison of results obtained from 67 clinical samples using existing RT-PCR, shell vial, cell culture and ELISA methods showed that QCM techniques were comparable in sensitivity and specificity to cell culture methods.

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Section: Introductionmentioning

confidence: 99%

The detection of influenza A and B viruses in clinical specimens using a quartz crystal microbalance

Hewa¹,

Tannock

Mainwaring³

et al. 2009

Journal of Virological Methods

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show abstract

“…Woo et al have shown clinical and financial benefits of rapid detection of respiratory viral infections in a pediatric population in Hong Kong, but no other studies address the clinical and financial impact of virology data (5). Doing et al described a technique of performing immunofluorescent testing (FA) on cytocentrifuged preparations which resulted in accurate, rapid detection of respiratory viral illnesses (4). In this study, we combined the methods used in these two studies to optimize rapid diagnosis and to evaluate its impact on patient care in a community teaching hospital in the United States.…”

mentioning

confidence: 99%

Clinical and Financial Benefits of Rapid Detection of Respiratory Viruses: an Outcomes Study

et al. 2000

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To assess the expected benefits of rapid reporting of respiratory viruses, we compared patients whose samples were processed using standard techniques such as enzyme immunoassays, shell vial assays, and culture tube assays (year 1) to patients whose samples were processed with the same standard techniques in addition to immunofluorescent testing (FA) directly on cytocentrifuged samples (year 2). The cytospin FA screened for influenza A and B viruses, respiratory syncytial virus (RSV), parainfluenza viruses 1 to 3, and adenovirus (DAKO Diagnostics Ltd.). The specificity of the cytospin FA for all viruses was 100%. The sensitivities for influenza A virus and RSV were 90 and 98%, respectively, but the sensitivities for influenza B virus and adenovirus were unacceptable (14.3 and 0%, respectively). However, since the former viruses account for >85% of our isolates from clinical specimens, the cytospin FA is an excellent screening test since the positive result was available within hours. The mean turnaround time for all positive viruses was 4.5 days in year 1 and 0.9 day in year 2 (P = 0.001). This rapid reporting resulted in physicians having access to information sooner, enabling more appropriate treatment. The mean length of stay in the hospital for inpatients with respiratory viral isolates was 10.6 days for year 1 versus 5.3 days for year 2. Mean variable costs for these patients was $7,893 in year 1 and $2,177 in year 2. After subtracting reagent costs and technological time, the savings in variable costs was $144,332/year. Summarizing, the cytospin FA markedly decreased turnaround time and was associated with decreased mortality, length of stay, and costs and with better antibiotic stewardship.

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“…Cytospin preparation of slides improves results. 15 For respiratory Advantages: Detect viruses before CPE is apparent, less expertise than conventional culture Disadvantages: Limited number of viruses, requires some expertise, requires cell culture facility and IF microscope, some BSL-3 or BSL-4 pathogens may be inadvertently grown in culture viruses, a pool of antibodies to several different pathogens can be used to screen a single cell spot for multiple viruses including 7 or 8 respiratory viruses (respiratory syncytial virus [RSV], influenza A and B, parainfluenza types 1, 2, 3, adenovirus, and human metapneumovirus). 16 For samples that screen positive, additional testing is required to identify infecting viruses.…”

Section: Viral Antigen Detectionmentioning

confidence: 99%

Rapid Diagnosis of Influenza

Peaper

Landry

2014

Clinics in Laboratory Medicine

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Much effort has been expended developing testing modalities for influenza viruses that are capable of providing rapid results to clinicians. Antigen-detection techniques, historically the only methods able to deliver results quickly, are still widely used despite concerns about sensitivity. Recently, nucleic acid amplification tests (NAATs), which can achieve rapid turnaround times and high sensitivity, have become available. In addition, NAATs can detect other respiratory pathogens. Although there are many theoretical advantages to rapid influenza testing, the clinical impact of testing in various patient populations must be considered against the cost and the analytical performance of the tests.

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Use of Fluorescent-Antibody Staining of Cytocentrifuge-Prepared Smears in Combination with Cell Culture for Direct Detection of Respiratory Viruses

Cited by 31 publications

References 13 publications

The detection of influenza A and B viruses in clinical specimens using a quartz crystal microbalance

The detection of influenza A and B viruses in clinical specimens using a quartz crystal microbalance

Clinical and Financial Benefits of Rapid Detection of Respiratory Viruses: an Outcomes Study

Rapid Diagnosis of Influenza

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