The assessment of a novel direct immunofluorescence technique as a rapid alternative to molecular techniques for the laboratory diagnosis of respiratory viral infection
Cardiff Metropolitan University
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Respiratory viruses are the most common cause of acute illness in otherwise healthy individuals, as well as causing significant morbidity and mortality in immunocompromised patients, those with underlying medical conditions or those at either end of the age scale. The epidemiology of different respiratory viruses can vary greatly, with regards to age group and timing of seasonal outbreaks [1, 2]. In recent years traditional laboratory testing methods for respiratory viruses, such as cell culture, have been replaced with more rapid and reliable molecular methods. However in busy periods, for example during winter respiratory virus season, large numbers of samples can lead to a delay in result reporting. The aim of this study was to determine the possibility of using a novel immunofluorescence kit as an alternative to molecular methods to provide more rapid results for specimens deemed to be urgent [3, 4]. 34 nasopharyngeal aspirates and 11 bronchial lavage specimens were simultaneously tested using the Diagnostic Hybrids D3 FastPoint L-DFA Respiratory Virus ID Kit and in-house real-time RT-PCR assay to detect Influenza A, Influenza B and RSV. 15 specimens were also tested using the Light Diagnostics Simulfluor DFA kit as an additional test where the FastPoint kit was unsuitable due to the viscosity of the samples. Considering PCR to be the Gold Standard, immunofluorescence correctly identified 26 of the 34 positive PCR samples. With 6 false negatives and 0 false positives this method proved to demonstrate 100% specificity and 81% sensitivity. 15 samples were repeated using the Simulfluor kit, with 2 of these deemed to contain insufficient cellular material for immunofluorescence testing. Although immunofluorescence testing is cheaper and less time consuming this study found the FastPoint kit to be unsuitable for considerably viscous samples and relatively subjective for samples which contained low amounts of cellular material. The small sample population of this study however may limit its reliability so further studies should be carried out as confirmation of these results.
BSc (Hons) Biomedical Science
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