Human Haemophilus Influenza B IgG ELISA Kit

Sample

Serum, plasma

Detection Method

iELISA

Intended Use

Enzyme immunoassay for the determination of IgG antibodies against polyribosylribitolphosphate (PRP) of Haemophilus influenzae type B in human serum and plasma.

Contents of Kit

1. 1 × 12 × 8, MTP. Microtiter Plate
Coated with PRP from Haemophilus influenzae Typ B. Ready to use!
2. 1 × 0.3 mL, ENZCONJ. CONC Conjugate, Concentrate
Anti-human-IgG-peroxidase; colored blue. Dilute before use!
3. 5 × 0.35 mL, CAL 1-5. Standard 1-5, Concentrate
Human sera with stabilizers and preservatives. Concentrations are lot specific as indicated on the bottle labels. Dilute before use!
4. 2 × 0.35 mL, POS LL, POS HL. Positive Control (Low and High), Concentrate
Positive control sera, LL, "Low Level", HL, "High Level"; for testing accuracy; human sera with stabilizers and preservatives.
Concentrations are lot specific as indicated on the bottle labels.
Dilute before use!
5. 2 × 75 mL, DILBUF. Incubation Buffer (Dilution Buffer)
0.01 M Tris/HCl; pH 7.4; contains detergent; 0.01 % (w/v) thimerosal; colored red. Ready to use!
6. 1 × 100 mL, WASHBUF CONC. Wash Buffer, Concentrate (10×)
Contains: phosphate buffer
7. 1 × 15 mL, TMB SUBS. TMB Substrate Solution
Substrate solution contains TMB (tetramethylbenzidine). Ready to use!
8. 1 ×15 mL, TMB STOP. TMB Stop Solution (0.5 M)
Stop solution, 0.5 M sulphuric acid. Ready to use!

Storage

The kit is shipped at ambient temperature and should be stored at 2 - 8°C. Keep away from heat or direct sunlight. The storage and stability of specimens and prepared reagents is stated in the corresponding chapters.
The microtiter strips are stable up to 6 months in the broken, but tightly closed bag when stored at 2 - 8°C.

Precision

Intraassay variation: 2 samples in the concentration range of the calibrators measured 20 times each with one lot in double detemination. The intraassay variation was ≤10%.
Interassay variation: POS HL and POS LL controls measured in 20 runs on different days with one lot in double determination. The interassay precision was between 9 and 12% respectively. Lower detection limit: ≤ 0.1μg/mL(lot-specific).

General Description

Haemophilus influenzae type B (HiB) is a very common cause of invasive critical infectious diseases in children up to the age of six. Following infection the symptoms of the disease include: Pericarditis, osteomyelitis, meningitis, encephalitis, pneumonia, sinusitis and otitis. In many cases the disease is lethal or leads to neurological damage, which cannot always be prevented by rapid antibiotic therapy. The underlying reason for the disease is very often a latent immunodeficiency with a specifically reduced humoral immune response to the polyribosylribitolphosphate (PRP) in the polysaccharide encapsulation of the bacterium. In children another reason is the immaturity of the immune system. Today often the term "immunocompromised patients" is used, comprising all acquired and innate specific and unspecific immunodeficiencies.
As a result, in children of 3 months of age or older a vaccination with different sorts of PRP-containing vaccines is recommended. This can lead to a clear reduction in the number of infections with Haemophilus influenzae type B.
The titer of antibodies produced by vaccination can be used to confirm whether the vaccination has been successful. The HiB IgG is used to measure the level of PRP-specific IgG-antibodies following a 4 - 6 weeks period after complete immunization to monitor the humoral immune status of children or other individuals at risk.
Monitoring of the humoral immunostatus after vaccination. Verification of the diagnosis Haemophilus influenzae type B infection by repeated monitoring of antibody concentrations. Risk assessment in immunocompromised patients leading to a failure of vaccination with a PRP-containing vaccine.
This group comprises:
Children under 2 years having had an infection with Haemophilus influenzae type B,
Children with chronic, recurring bacterial infections of the respiratory tract,
Children with chronic otitis,
Patients with confirmed humoral immuno-deficiencies (IgG-2-deficiency, IgA-deficiency),
Patients with confirmed granulocyte deficiencies,
Patients under chemo or cytostatic therapy,
Children after splenectomy,
Patients with sickle-cell anaemia,
Patients with trisomy 21 (Down) syndrome, and certain ethnic groups.