Validation of Single Radial Haemolysis assay: A reliable method to measure antibodies against influenza viruses

Trombetta, Claudia Maria; Perini, Daniele; Vitale, Licia; Stanzani, Valerio; Piccirella, Simona; Montomoli, Emanuele

doi:10.1016/j.jim.2015.04.009

Cited by 14 publications

(16 citation statements)

References 10 publications

(4 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The plates were stored in a humid box and incubated at 4°C for 16‐18 hours in the dark. After overnight incubation, the plates were incubated in a water bath at 37°C for 90 minutes, after which the diameters of the hemolysis areas were read in millimeters (mm) with a calibrated viewer …”

Section: Methodsmentioning

confidence: 99%

“…3,10,11 The advantages of the SRH assay are the small quantities of influenza virus and serum required, the ability to simultaneously analyze a large number of serum samples without pre-treatment (apart from complement inactivation) and unbiased results available after overnight incubation. 12 In addition, the assay detects small differences in antibody levels, distinguishes differentiates between closely related influenza strains, and is more sensitive for influenza B strains than the HI assay. [12][13][14][15] A hemolysis area of 25 mm 2 or greater is generally considered to be an immunological correlate of protection.…”

mentioning

confidence: 99%

“…12 In addition, the assay detects small differences in antibody levels, distinguishes differentiates between closely related influenza strains, and is more sensitive for influenza B strains than the HI assay. [12][13][14][15] A hemolysis area of 25 mm 2 or greater is generally considered to be an immunological correlate of protection. 16 Another widely used serological technique is the virus neutralization assay, which is recommended by the World Health Organization (WHO) for the measurement of functional antibodies against highly pathogenic avian viruses 6 and currently included in the EMA guidelines on influenza vaccines.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Comparison of hemagglutination inhibition, single radial hemolysis, virus neutralization assays, and ELISA to detect antibody levels against seasonal influenza viruses

Trombetta

Remarque

Mortier

et al. 2018

Influenza Resp Viruses

Self Cite

View full text Add to dashboard Cite

BackgroundThe immunological response to influenza vaccine and/or natural infection is evaluated by serological techniques, the most common being hemagglutination inhibition (HI), single radial hemolysis (SRH), and virus neutralization assays, which is commonly used in a micro‐neutralization (MN) format. ELISA is not officially required; however, this assay is able to measure different class‐specific antibodies. The four assays identify different sets or subsets of antibodies.ObjectivesThe aim of this study was to establish the correlation among four serological assays using four seasonal influenza strains.MethodsThe HI, SRH, MN assays, and ELISA were performed on four seasonal influenza strains.ResultsA strong positive correlation was found between HI and MN and between SRH and MN assays for influenza A strains. The B strains also showed good correlations among the three assays. A positive correlation was also found between ELISA and the “classical” assays for all strains. Concerning the correlates of protection, as defined by HI ≥ 40 and SRH ≥ 25 mm2, good agreement was observed for the influenza A strains. By contrast, the agreement for the B strains was very low.ConclusionsThere is a positive strong correlation among the four serological assays for both A and B strains, especially for the HI and MN assays. There is good agreement on correlates of protection between HI and SRH assays for the A strains, but very low agreement for the B strains, suggesting higher sensitivity of SRH than HI assay in detecting antibodies against the influenza B viruses.

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Comparison of hemagglutination inhibition, single radial hemolysis, virus neutralization assays, and ELISA to detect antibody levels against seasonal influenza viruses

Trombetta

Remarque

Mortier

et al. 2018

Influenza Resp Viruses

Self Cite

View full text Add to dashboard Cite

show abstract

“…The SRH assay enables antibodies to be quantified by measuring hemolysis areas [ 44 , 45 ], which are proportional to the antibody concentration, since they result from the binding between the antigen and the antibody. The assay is rapid, easily reproducible, and reliable, and also allows a high number of samples to be analyzed; for this reason, it is notably appropriate for large-scale studies, particularly epidemiological investigations [ 46 , 47 ].…”

Section: Evaluation Of Influenza Vaccine Efficacy: From Chmp Critementioning

confidence: 99%

Immunogenicity and Safety of the New Inactivated Quadrivalent Influenza Vaccine Vaxigrip Tetra: Preliminary Results in Children ≥6 Months and Older Adults

et al. 2018

Self Cite

View full text Add to dashboard Cite

Since the mid-1980s, two lineages of influenza B viruses have been distinguished. These can co-circulate, limiting the protection provided by inactivated trivalent influenza vaccines (TIVs). This has prompted efforts to formulate quadrivalent influenza vaccines (QIVs), to enhance protection against circulating influenza B viruses. This review describes the results obtained from seven phase III clinical trials evaluating the immunogenicity, safety, and lot-to-lot consistency of a new quadrivalent split-virion influenza vaccine (Vaxigrip Tetra®) formulated by adding a second B strain to the already licensed TIV. Since Vaxigrip Tetra was developed by means of a manufacturing process strictly related to that used for TIV, the data on the safety profile of TIV are considered supportive of that of Vaxigrip Tetra. The safety and immunogenicity of Vaxigrip Tetra were similar to those of the corresponding licensed TIV. Moreover, the new vaccine elicits a superior immune response towards the additional strain, without affecting immunogenicity towards the other three strains. Vaxigrip Tetra is well tolerated, has aroused no safety concerns, and is recommended for the active immunization of individuals aged ≥6 months. In addition, preliminary data confirm its immunogenicity and safety even in children aged 6–35 months and its immunogenicity in older subjects (aged 66–80 years).

show abstract

“…The assay is specifically suitable for use in large-scale clinical trials, owing to its rapidity, reproducibility and reliability. 22,23 The SRH assay has a higher sensitivity for influenza B strains. 24,25 The HI assay is the gold-standard assay of antibody titers against HA, and is based on erythrocyte agglutination due to the ability of antibodies that specifically recognize HA to inhibit the binding of viral surface protein HA to sialic acid sites on the surface of red blood cells.…”

Section: Introductionmentioning

confidence: 99%

The use of cell-mediated immunity for the evaluation of influenza vaccines: an upcoming necessity

Gianchecchi

Torelli

Montomoli

2019

Human Vaccines & Immunotherapeutics

View full text Add to dashboard Cite

Influenza vaccines are a fundamental tool for preventing the disease and reducing its consequences, particularly in specific high-risk groups. In order to be licensed, influenza vaccines have to meet strict criteria established by European Medicines Agency. Although the licensure of influenza vaccines started 65 years ago, Hemagglutination Inhibition and Single Radial Hemolysis are the only serological assays that can ascertain correlates of protection. However, they present evident limitations. The present review focuses on the evaluation of cell-mediated immunity (CMI), which plays an important role in the host immune response in protecting against virus-related illness and in the establishment of longterm immunological memory. Although correlates of protection are not currently available for CMI, it would be advisable to investigate this kind of immunological response for the evaluation of nextgeneration vaccines.

show abstract

Validation of Single Radial Haemolysis assay: A reliable method to measure antibodies against influenza viruses

Cited by 14 publications

References 10 publications

Comparison of hemagglutination inhibition, single radial hemolysis, virus neutralization assays, and ELISA to detect antibody levels against seasonal influenza viruses

Comparison of hemagglutination inhibition, single radial hemolysis, virus neutralization assays, and ELISA to detect antibody levels against seasonal influenza viruses

Immunogenicity and Safety of the New Inactivated Quadrivalent Influenza Vaccine Vaxigrip Tetra: Preliminary Results in Children ≥6 Months and Older Adults

The use of cell-mediated immunity for the evaluation of influenza vaccines: an upcoming necessity

Contact Info

Product

Resources

About