Influenza A IgG ELISA Kit

Name: Influenza A IgG ELISA Kit
SKU: DEIA353
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Human serum, plasma, or cerebrospinal fluid

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The Influenza A IgG ELISA Kit is a qualitative and quantitative immunoassay for the detection of specific human IgG antibodies directed against the nucleo- and matrix proteins of Influenza A virus in serum, plasma or cerebrospinal fluid. The test supports the diagnosis of an existing or past infection. The test allows the detection of intrathecally formed IgG antibodies for CSF diagnostics. The test system can be processed manually or automatically. The tests are designed for professional use. An application for screening procedures in the context of transplantation or transfusion is excluded.

Contents of Kit

Test Components	Description	Pieces/Volume
Break apart microtiter test strips, each with eight antigen coated single wells (altogether 96) MTP	1 frame; the coating material is inactivated	12 pieces
Standard serum (ready-to-use) STD	Human serum in protein containing phosphate buffer; negative for anti-HIV Ab, HBs-Ag (Hepatitis B-Virus surface antigen) and anti-HCV Ab; preservative: < 0.1 % sodium azide; colouring: Amaranth O	2 x 2 mL
Negative control serum (ready-to-use) NEG	Human serum in protein containing phosphate buffer; negative for anti-HIV Ab, HBs-Ag (Hepatitis B-Virus surface antigen) and anti-HCV Ab; preservative: < 0.1 % sodium azide; colouring: Lissamin Green V	2 mL
Anti-human IgG conjugate (ready-to-use) APC	Anti-human IgG polyclonal antibody, conjugated to alkaline phosphatase, stabilised with protein stabilisation solution; preservative: < 0.1 % methylisothiazolone, < 0.1 % bromnitrodioxane	13 mL
Washing solution concentrate (sufficient for 1000 mL) WASH	Sodium chloride solution with Tween 20 and 30 mM Tris/HCl, pH 7.4; preservative: < 0.1 % sodium azide	33.3 mL
Dilution buffer (ready-to-use) DILB	Protein containing phosphate buffer with Tween 20; preservative: < 0.1 % sodium azide; colouring: 0.01 g/L Bromphenol blue	2 x 50 mL
Stopping solution (ready-to-use) STOP	< 0.1 N sodium hydroxide, 40 mM EDTA	15 mL
Substrate (ready-to-use) pNPP	Para-nitrophenylphosphate in solvent free buffer; preservative: < 0.1 % sodium azide	13 mL
Quality control certificate with standard curve and evaluation table INFO	Quantification of antibodies in U/mL	2 pages

Storage

2 - 8 °C

Performance Characteristics

Sensitivity and specificity: The Influenza A Virus IgG test was evaluated by the analysis of 68 serum samples from patients and 105 samples from blood donors in comparison to CFT, with the presumption that IgA antibody production after an infection and the synthesis of complement binding antibodies takes place simultaneously. Sera classified as borderline were not included in the calculation.

Test	Sensitivity	Specificity
Influenza A Virus IgG	92.3 %	90.1 %

Reproducibility: see Precision.
Cross-reactivities: Positive sera (10 sera each) for Influenza B Virus IgG, Bordetella pertussis IgG, Helicobacter pylori IgG, Respiratory Syncytial Virus IgG, Adenovirus IgG, Parainfluenza Virus IgG and Mycoplasma pneumoniae IgG have been tested as well as sera positive for rheumatoid factor (RF) and anti-nuclear antibodies (ANA). Within this internal evaluation, potential cross-reactivities with one Influenza B Virus IgG, two Respiratory Syncytial Virus IgG, one RF and two ANA positive serum samples have been observed. All reactivities have been confirmed by positive or borderline results in the reference assay. Other cross-reactivities cannot be ruled out in general.
Interfering substances: No interferences have been detected for sera with concentrations up to 2.00 g/L hemoglobin, 11.50 g/L lipemia/triglyceride or 0.201 g/L bilirubin (conjugated and unconjugated). Other interfering substances cannot be ruled out in general.
Hook effect: Evaluations show that the test exhibits no hook effect.

Precision

Reproducibility (Influenza A Virus IgG):

Sample	Mean Value (OD)	Repeatability CV (%)	Mean Value (OD)	Reproducibility CV (%)
Serum 1	0.160	9.2	0.159	7.2
Serum 2	0.399	4.4	0.435	8.2
Serum 3	1.219	3.9	1.451	4.3

Detection Range

2 - 200 U/mL

Detection Limit

2 U/mL (Limit of Quantification)

Sensitivity

Diagnostic sensitivity: 92.3 %; Limit of Quantification: 2 U/mL

General Description

The orthomyxovirus Influenza A has a worldwide distribution with natural reservoirs including humans, as well as a wide range of mammals and birds. The virus has two important components, the haemagglutinin and neuraminidase proteins, which are embedded in the virus envelope. These are responsible for the characteristic high immunogenic variability of the virus resulting from the high mutation rate of RNA and the potential for reassortment of the segmented genome.
Influenza is an infection of the respiratory tract. Transmission of the highly infectious Influenza A virus is primarily by aerosol. The incubation period ranges from a few hours to several days. The spectrum of disease is very variable and ranges from asymptomatic to death due to influenza induced pneumonia. A rapid onset of symptoms is characteristic with fever, cough, headache and muscle pains appearing within hours.
In the routine diagnostic laboratory, PCR serves to demonstrate the presence of Influenza A virus in the early stages of disease, while ELISA is a reliable method for the demonstration of pathogen-specific antibodies. The use of conserved nucleo- (NP) or matrix proteins (M) as antigens permits the detection of antibodies independent of the infecting Influenza A virus subtype, and the detected antibodies generally persist only for weeks or months post infection, allowing better differentiation between past and acute infections.

Standard Curve

A lot-specific standard curve (4-parameter logistic function) together with an evaluation table is provided on the quality control certificate included in each test kit. The obtained OD values are assigned to the corresponding antibody activities (U/mL). The substrate blank must be subtracted from all OD values prior to evaluation; the mean OD value of the standard serum (tested in duplicate) has to be used.

Reconstitution And Storage

Bring all reagents to room temperature before testing.

Reagent	Storage	Stability
Microtiter strips (coated with antigen)	unopened at 2 - 8 °C; after opening at 2 - 8 °C in closed aluminum bag with desiccant	see expiry date; 6 months after opening
Control sera / Standard sera	unopened at 2 - 8 °C; after opening at 2 - 8 °C	see expiry date; 6 months after opening
Conjugate	unopened at 2 - 8 °C; after opening at 2 - 8 °C	see expiry date; 6 months after opening
Dilution buffer	unopened at 2 - 8 °C; after opening at 2 - 8 °C	see expiry date; 6 months after opening
Washing solution	unopened / after opening at 2 - 8 °C; working dilution at 2 - 8 °C; working dilution at room temperature	see expiry date; 2 weeks (working dilution at 2 - 8 °C); 1 week (working dilution at room temperature)
Substrate	unopened at 2 - 8 °C; after opening at 2 - 8 °C	see expiry date; 6 months after opening
Stopping solution	unopened at 2 - 8 °C; after opening at 2 - 8 °C	see expiry date; 6 months after opening

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

Infectious Disease Test Kits and Positive Controls

QA & Reviews

Customer Reviews

References

Targeting the endolysosomal host-SARS-CoV-2 interface by clinically licensed functional inhibitors of acid sphingomyelinase (FIASMA) including the antidepressant fluoxetine

EMERGING MICROBES & INFECTIONS

Authors: Schloer, Sebastian; Brunotte, Linda; Goretzko, Jonas; Mecate-Zambrano, Angeles; Korthals, Nadia; Gerke, Volker; Ludwig, Stephan; Rescher, Ursula

Abstract

The Coronavirus Disease 2019 (COVID-19) pandemic caused by the Severe Acute Respiratory Syndrome Related Coronavirus 2 (SARS-CoV-2) is a global health emergency. As only very limited therapeutic options are clinically available, there is an urgent need for the rapid development of safe, effective, and globally available pharmaceuticals that inhibit SARS-CoV-2 entry and ameliorate COVID-19 severity. In this study, we explored the use of small compounds acting on the homeostasis of the endolysosomal host-pathogen interface, to fight SARS-CoV-2 infection. We find that fluoxetine, a widely used antidepressant and a functional inhibitor of acid sphingomyelinase (FIASMA), efficiently inhibited the entry and propagation of SARS-CoV-2 in the cell culture model without cytotoxic effects and also exerted potent antiviral activity against two currently circulating influenza A virus subtypes, an effect which was also observed upon treatment with the FIASMAs amiodarone and imipramine. Mechanistically, fluoxetine induced both impaired endolysosomal acidification and the accumulation of cholesterol within the endosomes. As the FIASMA group consists of a large number of small compounds that are well-tolerated and widely used for a broad range of clinical applications, exploring these licensed pharmaceuticals may offer a variety of promising antivirals for host-directed therapy to counteract enveloped viruses, including SARS-CoV-2.

Influenza A virus infection induces liver injury in mice

MICROBIAL PATHOGENESIS

Authors: Zhang, Shouping; Hu, Bin; Xu, Jingfei; Ren, Qiuxuan; Wang, Lirong; Wang, Sanhu

Abstract

Respiratory infections such as SARS-CoV in humans are often accompanied by mild and self-limiting hepatitis. As a respiratory disease, influenza A virus (IAV) infection can lead to hepatitis, but the mechanism remains unclear. This study aimed to investigate the occurrence of hepatitis by establishing a model for infected mice for three different subtypes of respiratory IAVs (H1N1, H5N1, and H7N2). Histological analysis was performed, and results showed increase serum aminotransferase (ALT and AST) levels and evident liver injury on days 3 and 7, especially on day 5 post infection. Immunohistochemistry (IHC) results indicated a wide distribution of IAV's positive signals in the liver of infected mice. Real-time PCR results further revealed a similar viral titer to IHC that presented a remarkedly positive correlation with histology injury. All these data showed that the mouse model suitably contributed valuable information about the mechanism underlying the occurrence of hepatitis induced by respiratory influenza virus.