Subtyping Influenza A Virus with Monoclonal Antibodies and an Indirect Immunofluorescence Assay

Johnson, Jennifer G.; Higgins, Angela D.; Navarro, Adriana; Huang, Yung T.; Esper, Frank L.; Barton, Nicole; Esch, D Aniela W; Shaw, Carl; Olivo, Paul D.; Miao, Lynn Yihong

doi:10.1128/jcm.01237-11

Cited by 13 publications

(8 citation statements)

References 15 publications

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“…20 By changing the specific antibodies used, this method can be modified to detect all types of influenza viruses. 21 To improve the accuracy of virus titration, we tried to control the activity of HA and NA by optimizing the culture media. Offspring influenza virus particles produced during the course of the ELISPOT titration assay may infect neighboring cells to form extra spots, resulting in a decrease in accuracy.…”

Section: Discussionmentioning

confidence: 99%

Establishment of a rapid ELISPOT assay for influenza virus titration and neutralizing antibody detection

Wang

Huang

Hong

et al. 2020

Journal of Medical Virology

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Seasonal influenza is an acute respiratory infection causing around 500 000 global deaths annually. There is an unmet medical need to develop more effective antiviral drugs and vaccines against influenza infection. A rapid, accurate, high‐throughput titration assay for influenza virus particles or neutralizing antibodies would be extremely useful in these research fields. However, commonly used methods such as tissue culture infective dose and plaque‐forming units (PFU) for virus particle quantification, and the plaque reduction neutralization test (PRNT) for antibody determination are time‐consuming, laborious, and have limited accuracy. In this study, we developed an efficient assay based on the enzyme‐linked immunospot (ELISPOT) technique for the influenza virus and neutralizing antibody titration. Two broad‐spectrum antibodies recognizing the nucleoproteins of influenza A and B viruses were used in the assay to broadly and highly sensitively detect influenza virus‐infected cells at 16 hours postinfection. An optimized cell culture medium with no tosyl phenylalanyl chloromethyl ketone trypsin and high dose oseltamivir acid was used to improve quantitation accuracy. This ELISPOT assay displayed a good correlation (R2 = 0.9851) with the PFU assay when used to titrate 30 influenza virus isolates. The assay was also applied to measure influenza‐neutralizing antibodies in 40 human sera samples, showing a good correlation (R2 = 0.9965) with the PRNT assay. This ELISPOT titration assay is a rapid, accurate, high‐throughput assay for quantification of influenza virus and neutralizing antibodies, and provides a powerful tool for research into and development of drugs and vaccines against influenza.

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Section: Discussionmentioning

confidence: 99%

Establishment of a rapid ELISPOT assay for influenza virus titration and neutralizing antibody detection

Wang

Huang

Hong

et al. 2020

Journal of Medical Virology

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“…IFA was also used for direct detection of HSV antigen in clinical specimens with sensitivity of 84.6% and specificity of 95.7%, compared to the tissue culture method [ 151 ]. Moreover, IFA was applied for subtyping of influenza A virus with 100% agreement to RT-PCR [ 153 ]. IFA was also used for the detection of EBV [ 21 ] and as a confirmatory test for HIV-1 [ 154 ].…”

Section: Immunological Diagnostic Techniques Of Medical Virusesmentioning

confidence: 99%

“…HI assay is laborious and time-consuming. The interpretation of the assay results between laboratories may be different as no standard reagents are available for the assay [ 153 , 157 , 166 ]. In case of IF assay, prolonged exposure of specimen to UV light leads to fading of fluorescence that could result in false-negative test [ 167 ].…”

Section: Immunological Diagnostic Techniques Of Medical Virusesmentioning

confidence: 99%

Molecular and Immunological Diagnostic Techniques of Medical Viruses

Reta

Tessema

Ashenef

et al. 2020

International Journal of Microbiology

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Viral infections are causing serious problems in human population worldwide. The recent outbreak of coronavirus disease 2019 caused by SARS-CoV-2 is a perfect example how viral infection could pose a great threat to global public health and economic sectors. Therefore, the first step in combating viral pathogens is to get a timely and accurate diagnosis. Early and accurate detection of the viral presence in patient sample is crucial for appropriate treatment, control, and prevention of epidemics. Here, we summarize some of the molecular and immunological diagnostic approaches available for the detection of viral infections of humans. Molecular diagnostic techniques provide rapid viral detection in patient sample. They are also relatively inexpensive and highly sensitive and specific diagnostic methods. Immunological-based techniques have been extensively utilized for the detection and epidemiological studies of human viral infections. They can detect antiviral antibodies or viral antigens in clinical samples. There are several commercially available molecular and immunological diagnostic kits that facilitate the use of these methods in the majority of clinical laboratories worldwide. In developing countries including Ethiopia where most of viral infections are endemic, exposure to improved or new methods is highly limited as these methods are very costly to use and also require technical skills. Since researchers and clinicians in all corners of the globe are working hard, it is hoped that in the near future, they will develop good quality tests that can be accessible in low-income countries.

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“…Due to the complexity of the assays, the variations in detection sensitivity and specificity are probably caused by differences in technical expertise, specimen type, and quality (the presence of an adequate number of infected cells), as well as patient selection [27,47,48]. Even with the variation in detection sensitivity and their relatively extensive infrastructure requirements, the immunofluorescence staining methods have demonstrated high accuracy in detecting influenza virus and have been adapted for identification of specific influenza subtypes using monoclonal antibodies specific for HA subtypes, such as H1, H3, H5, and H7 [49][50][51]. DFA and IFA are valuable in confirming rapid diagnostic testing results [31,[52][53][54].…”

Section: Current Detection Technologiesmentioning

confidence: 99%

Evolving Gene Targets and Technology in Influenza Detection

Malanoski

Lin

2013

Mol Diagn Ther

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Influenza viruses cause recurring epidemic outbreaks every year associated with high morbidity and mortality. Despite extensive research and surveillance efforts to control influenza outbreaks, the primary mitigation treatment for influenza is the development of yearly vaccine mixes targeted for the most prevalent virus strains. Consequently, the focus of many detection technologies has evolved toward accurate identification of subtype and understanding the evolution and molecular determinants of novel and pathogenic forms of influenza. The recent availability of potential antiviral treatments are only effective if rapid and accurate diagnostic tests for influenza epidemic management are available; thus, early detection of influenza infection is still important for prevention, containment, patient management, and infection control. This review discusses the current and emerging technologies for detection and strain identification of influenza virus and their specific gene targets, as well as their implications in patient management.

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Subtyping Influenza A Virus with Monoclonal Antibodies and an Indirect Immunofluorescence Assay

Cited by 13 publications

References 15 publications

Establishment of a rapid ELISPOT assay for influenza virus titration and neutralizing antibody detection

Establishment of a rapid ELISPOT assay for influenza virus titration and neutralizing antibody detection

Molecular and Immunological Diagnostic Techniques of Medical Viruses

Evolving Gene Targets and Technology in Influenza Detection

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