Campylobacter IgG ELISA Kit

Sample

Serum, plasma

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Campylobacter IgG kit is a qualitative or quantitative in vitro test for the detection and safe identification of IgG antibodies against Campylobacter jejuni and Campylobacter coli in human serum or plasma. The Campylobacter IgG kit is a screening test based on the principle of an indirect sandwich ELISA.

Contents of Kit

1. Wash buffer (10× the concentration) Contains phosphate buffer, NaCl, detergent and preservatives MIT (0,01%) and Oxypyrion (0,1%), 100 mL
2. Dilution buffer (ready-for-use) Contains protein, detergent and blue dye, preservatives MIT (0,01%) and Oxypyrion (0,1%), 125 mL
3. Chromogenic substrate tetramethylbenzidine (TMB, ready-for-use), 12 mL
4. Stop solution 24,9 % Phosphoric acid (H₃PO₄) , 12 mL
5. Microplate (section marked in blue) coated with recombinant Campylobacter antigens in vacuum-pressure sealed bag, 12×8 wells
6. Positive control (brown screw cap) Preservatives: MIT (0,1%) and Oxypyrion (0,1). 450μL
7. Cutoff control (orange screw cap) Preservatives: MIT (0,1%) and Oxypyrion (0,1). 450μL
8. Negative control (white screw cap) Preservatives: MIT (0,1%) and Oxypyrion (0,1). 450μL
9. Anti-human IgA conjugate (101 times the concentration, blue screw cap) contains NaN₃ (<0,1%), MIT (<0,01%), Chloracetamide (<0,1%). 500 μL
10. Instructions for use 1
11. Evaluation form 1
12. Sealing tape 2 pieces

Storage

2°C - 8°C

Performance Characteristics

1. Intra-Assay-Variance:
A serum was examined on 48 cavities of a microtiterplate. The variation coefficient (VC) was calculated:
VC (IgG) = 4,6%, VC (IgA) = 4,1%.
2. Inter-Assay-Variance:
7 sera of different ODs ware examined in 8 different determinations. The variation coefficient was
calculated for each single serum (U/ml):
VC (IgG) < 12%, VC (IgA) < 12 %

General Description

The genus Campylobacter comprises gram-negative, spiral-shaped, microaerophilic, mesophilic to thermophilic bacteria with bipolar flagella. In 1963, Sebald and Vernon named a bacterium Escherich
had described as early as 1889 Campylobacter jejuni. Isolation from stool samples was achieved in 1972 by Dekeyser et al. Jones et al. made out a food association in 1981/1982. In taxonomic terms, Campylobacter is classified with the epsilon subdivision of the Proteobacteria.
The pathogen reservoir is mainly the intestinal tract of warm-blooded wild, domestic and pet animals (birds and mammals). Intestinal Campylobacter infections are the second most frequent enteric bacterial infections reported in Germany after enteric salmonellosis (Robert Koch Institute 2004: RKI Infektionsepidemiologisches Jahrbuch meldepflichtiger Krankheiten 2006: Salmonella 52.319, Campylobacter 51.764), whereby Campylobacter jejuni is much more frequent, accounting for over 90% of cases as compared to Campylobacter coli at approx. 9%.
Contaminated foods (mainly poultry) and drinking water (tropical countries) constitute the main sources of infection. Unreported cases not reflected in the statistics probably outnumber reported cases many times over. Annual incidence rates vary seasonally (higher rates in the summer months) as well as regionally, with diagnoses following travel to warm countries throughout the year due to considerably higher incidence in Third World countries.
Acute disease
The course of infection with Campylobacter is almost exclusively oral, the pathogen is highly adapted to the intestinal tract. Systemic infections up to and including meningitis are observed less frequently (especially in immunocompromised patients, infants and the elderly) (Skirrow & Blaser, 2000). The incubation period is brief (1-7 days) and presumably depends on the infective dose. Besides nearly asymptomatic (clinically inapparent) courses, infected persons suffer from painful gastrointestinal symptoms with sometimes bloody diarrhoea, fever, meningism and myalgias. The acute clinical picture persists for only a few days in most patients. The course of the infection is self-limiting in most cases, although severe losses of fluids and electrolytes must be replaced as required.
Sequelae:
Reactive arthritis (ReA) is among the known sequelae to enteral, and frequently urogenital infections as well caused by certain bacterial and viral pathogens, with onset a few weeks after the primary infection. In addition to urogenital mycoplasmas and chlamydiae, enteral salmonellae, shigellae and yersiniae, Campylobacter spp. are prominent among secondary pathogen diagnoses (Locht & Krogfeldt, 2002; Cox et al., 2003; Hannu et al., 2004). The background pathophysiological mechanism is presumably molecular mimicry of cross-reacting antibodies to antigens of the synovial membrane. Some cases of arthritis of unclear genesis can be correctly diagnosed as post-infection Reactive Arthritis, and not rheumatoid arthritis, on the basis of the appropriate laboratory parameters.
Similarly to Reactive Arthritis, Guillain-Barré-Syndrom (GBS) may also develop a few weeks after infections, usually enteral or respiratory, caused by certain bacterial and viral pathogens. The presumed level of incidence is between 1:1,000 and 1:10,000. GBS is an acute, immunomediated polyradicular neuropathy caused by an abnormal humoral immune response to the peripheral myelin sheath and/or the neural axon. A number of different viruses (e.g. CMV, EBV, VZV, measles virus, mumps virus), Mycoplasma pneumoniae and bacteria (Borrelia burgdorferi, Haemophilus influenzae) can cause a GBS, but various case-control studies have implicated that mainly Campylobacter spp., and in particular Campylobacter jejuni (Nachamkin et al., 2000; Prendergast et al., 2004; Gilbert et al., 2004; Leonard et al., 2004) as prior agent with a frequency level of 30%, based mainly on serological finding. Direct pathogen detection methods produced negative results in most cases. In pathophysiological terms, pathogen-specific antibodies cross-react with neuronal antigens (e.g. Moran et al., 2000; Yuki et al., 2004), thereby inducing inflammation mediators to invade macrophages, resulting in subsequent local micro-oedemas with transitory or permanent failure of the affected neurones. Neurological failures in GBS therefore include purely motor ("AMAN" = acute motor axonal neuropathy, "AIDP" = acute inflammatory demyelinating polyneuropathy) as well as sensory and mixed types ("ANSAM" = acute motor-sensory axonal neuropathy). Mild and reversible courses are observed as well as severe courses with permanent paralysis. About 5% of GBS cases are terminal. The course of disease appears to be directly proportional to the specific antibody titre. Clinical observations and initial findings from surveillance studies on "flaccid paralysis" (differential diagnosis: poliomyelitis) suggest that, in pathophysiological terms, discrete, transitory paralysis symptoms that take the form of signs of exhaustion in the extremities correspond much more frequently to an inapparent GBS than has been assumed to date.