Bordetella pertussis IgM ELISA Kit

Sample

Serum, plasma

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The Bordetella pertussis IgM is quantitative and qualitative test for the detection of human antibodies in serum or plasma against Bordetella pertussis. For Research Use Only. Not for use in diagnostic procedures.

Contents of Kit

1. Break apart microtiter test strips each with 8 antigen coated single wells (altogether 96) MTP, 1 frame, the coating material is inactivated.
2. Standard serum (ready-to-use) STD, 2 × 2 ml. Human serum in phosphate buffer with protein; negative for anti-HIV-Ab, anti-HBs-Ag (Hepatitis B-Virus-surface antigen) and anti-HCV-Ab; preservative: < 0.1 % sodium azide coloring: Amaranth O.
3. Negative control serum (ready-to-use) NEG, 2 ml. Human serum in phosphate buffer with protein; negative for anti-HIV, anti-HBs (Hepatitis B-Virus-surface antigen) and anti-HCV; preservative: < 0.1 % sodium azide coloring: Lissamin green V.
4. Anti-human-IgM-conjugate (ready-to-use) APC, 13 ml. Anti-human-IgM from goat (polyclonal), conjugated to alkaline phosphatase, stabilized with protein stabilization solution preservative: 0.01 % methylisothiazolone, 0.01 % bromnitrodioxane.
5. Washing solution concentrate (sufficient for 1 litre) WASH, 33.3 ml. Sodium chloride solution with Tween 20, 30 mM Tris preservative: < 0.1 % sodium azide.
6. Dilution buffer DILB, 2 × 50ml. Phosphate buffer with protein and Tween 20; preservative: < 0.1 % sodium azide 0.01 g/l Bromphenol blue sodium salt.
7. Stopping solution STOP, 15 ml. 1.2 N sodium hydroxide.
8. Substrate (ready-to-use) pNPP, 13 ml. Para-nitrophenylphosphate, solvent free buffer preservative: < 0.1 % sodium azide (Substrate in unopened bottle may have a slightly yellow color. This does not reduce the quality of the product!)
9. Quality control certificate with standard curve and evaluation table INFO, 1. (quantification of antibodies in IU/ml or U/ml)

Storage

1. Microtiter strips (antigen). Unopened (see expiry date on microtiter plate). After opening at 2-8°C in closed aluminum bag with desiccant (minimum shelf-life 4 weeks). Strips which are not used must be stored in the press-seal bag of aluminum compound foil under dry and airtight conditions! Shelf-life in case of proper use and storage until expiry date.
2. Control sera / standard sera. After opening at 2-8°C (until expiry date, 24 months from date of production)
3. Conjugate. Ready-to-use solution, at 2-8°C. Avoid contamination (sterile tips!) (until expiry date, 28 months from date of production)
4. Dilution buffer. After opening at 2-8°C. Discard cloudy solutions! (24 months). Unopened (until expiry date; 36 months from date of production)
5. Washing solution. Concentrate after opening at 2-8°C (until expiry date). Working dilution at 2-8°C (2 weeks). Working dilution at room temperature (1 week). Bottles used for the working dilution should be cleaned regularly, discard cloudy solutions.
6. Substrate. ready-to-use solution at 2-8°C, protected from light! (until expiry date, 24 months from date of production). Avoid contamination (sterile tips!) Discard when solution turns yellow (extinction against distilled water > 0.25).
7. Stopping solution. After opening at room temperature (until expiry date).

General Description

Four species, Bordetella pertussis, B. parapertussis, B. bronchiseptica and B. avium belong to the bacterial genus of Bordetella. These bacteria are ubiquitous, small (0.2 – 0.5 μm), gram-negative obligatory aerobic coccoid single or paired bacteria. The ideal temperature for reproduction is 35 - 37 °C. B. pertussis and B parapertussis are human pathogens causing whooping cough (pertussis) and a milder form, which is less common (atypical pertussis). The other two species mostly cause respiratory diseases in warm-blooded animals. However, in rare cases (e.g. close contact with animals) B. bronchiseptica is an opportunistic secondary human pathogen in case of infections like bronchitis, pneumonia or wound infections. Even though there are phenotypic differences between the three human pathogenic Bordetella species, their DNA is 72-94% homologous. Therefore, it is currently being discussed whether to combine them in a single species or three subspecies of one species.
Bordetella pertussis causes whooping cough, a worldwide infectious disease that is transmitted from person to person by droplet infection. Especially children at the age of 0-4 years are affected, and the mortality of infected infants is high.
The attachment of B. pertussis to the tuft of ciliated cells in the mucosa of the human respiratory tract is mediated by adhesins. An important adhesion protein and an equally important immunogen is the so called filamentous hemagglutinin (FHA). Colonizing of the respiratory tract and establishment of infection are facilitated by the synergistic action of several virulence factors.
A significant virulence factor is pertussis toxin (PT), which is exclusively synthesized and secreted by B. pertussis. Multiple biologic effects like leukocytosis, lymphocytosis, mitogenicity and increased sensitivity to histamine are caused by PT.
The typical whooping cough can be divided into three stages: the catarrhal phase, the convulsivum (paroxysmal) phase, and the decrementi phase.
The ELISA technique is the most commonly chosen method for B. pertussis antibody detection.
A whole-cell antigen preparation of B. pertussis is used in the Bordetella pertussis IgM ELISA.