Cocci Ab ELISA Kit

Name: Cocci Ab ELISA Kit
SKU: DEIABL89
Rating: 5 (1 reviews)

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

COA

SDS

Datasheet

Summary

Sample

serum and cerebrospinal fluid (CSF)

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The clarus Coccidioides Antibody Enzyme Immunoassay (EIA) is used for the qualitative detection of IgM and IgG antibodies directed against TP and CF antigens from Coccidioides species as an aid in the diagnosis of coccidioidomycosis in serum and cerebrospinal fluid (CSF).

Contents of Kit

1. 10× Specimen Diluent: 20 mL Concentrated, buffered protein solution with a preservative.
2. 20× Wash Buffer: 50 mL Concentrated wash buffer with a preservative.
3. CF Antigen-Coated Microwells: 96 Color-coded (blue = CF) microwell plate featuring breakaway polystyrene microwells.
4. TP Antigen-Coated Microwells: 96 Color-coded (clear = TP) microwell plate featuring breakaway polystyrene microwells.
5. CF Calibrator Cutoff: 1.5 mL × 2 Anti-Coccidioides CF antibodies in a buffered protein solution (with a preservative) for establishing the cutoff signal for calculating CF EIA units.
6. TP Calibrator Cutoff: 1.5 mL × 2 Anti-Coccidioides TP antibodies in a buffered protein solution (with a preservative) for establishing the cutoff signal for calculating TP EIA units.
7. CF Enzyme Conjugate: 10 mL Afffnity-puriffed rabbit anti-human IgG antibodies conjugated to horseradish peroxidase (HRP) in a buffered protein solution with a preservative.
8. TP Enzyme Conjugate: 10 mL Afffnity-puriffed rabbit anti-human IgM antibodies conjugated to horseradish peroxidase (HRP) in a buffered protein solution with a preservative.
9. TMB Substrate : 20 mL Buffered solution containing urea peroxide and tetramethylbenzidine. TMB substrate is light-sensitive and should be kept out of direct light.
10. Stop Solution: 20 mL 2 N sulfuric acid. CAUTION: AVOID CONTACT WITH SKIN. FLUSH WITH WATER IF CONTACT OCCURS.
+. Positive Control: 1.5 mL × 2 Anti-Coccidioides antibodies in a buffered protein solution containing a preservative.

Storage

1. The entire test kit should be stored at 2-8 °C until the expiration dates listed on the labels. All reagents not used during testing should be returned to 2-8 °C storage promptly after use.
2. Unused microwells (3 and 4 in Reagents And Materials Provided) should be placed in the resealable Mylar bags and sealed immediately after opening and stored at 2-8 °C. Care should be taken to ensure the desiccant pouch remains in the bag with unused microwells.

Performance Characteristics

SERUM:
PERCENT AGREEMENT TO COMPLEMENT FIXATION/IMMUNODIFFUSION (N=1696)
The positive and negative percent agreement of the clarus Coccidioides Antibody EIA was evaluated versus Coccidioides complement fixation (CF). Discrepant results (EIA+/CF- and EIA-/CF+) were then tested using immunodiffusion (ID) for majority consensus. A total of 1696 serum samples were tested on the clarus Coccidioides Antibody EIA at a 1:441 dilution, using a cut-off value of 1.5 EIA Units.
Note: Positive results on either IgM and/or IgG plates indicates a positive result for the specimen.

CSF:
SENSITIVITY AND SPECIFICITY TO EORTC/MSG CLINICAL CRITERIA (N=81)
The sensitivity and specificity of the clarus Coccidioides Antibody EIA were evaluated on patients classified as having proven coccidioidomycosis according to the EORTC/MSG consensus definitions.A total of 81 CSF samples were tested on the clarus Coccidioides Antibody EIA at a 1:21 dilution, using a cut-off value of 0.3 EIA Units. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated using this data.
Note: Positive results on either IgM and/or IgG plates indicates a positive result for the specimen.

General Description

Coccidioidomycosis is an airborne, infectious disease that is caused by the Coccidioides spp. organisms. Coccidioides are dimorphic fungi that thrive in arid desert soils and environments with mild winters and dry summers. Exposure occurs when these microscopic spores are inhaled into the lungs. Infection can lead to respiratory diseases, and occasionally, diseases affecting other systems. Though endemic in the southwestern United States and Mexico, increased travel to the endemic areas has also increased the incidence in non-endemic areas.
Coccidioidomycosis should be considered whenever patients display flu-like symptoms and have lived or traveled to the endemic areas. Coccidioidomycosis presents a diagnostic challenge to the physician and laboratorian. The symptoms of most early coccidioidal infections substantially mimic those of other respiratory infections. In addition, the organisms can be difffcult to demonstrate via culture and histologically, even after repeated attempts. Therefore, speciffc laboratory testing is usually required to establish a diagnosis of coccidioidomycosis. Serologic tests have served for several decades as aids in the diagnosis and management of coccidioidomycosis.
Complement ffxation (CF), immunodiffusion (ID), and latex agglutination (LA) have been the most commonly used serologic methods. The CF assay is sensitive; however, its performance is complex and labor-intensive. Additionally, the CF assay exhibits low speciffcity due to cross-reactive antibodies that recognize carbohydrate moieties common to several fungi. The ID assay is more speciffc but less sensitive than the CF assay. Additionally, the ID assay takes 48 hours to perform and requires highly skilled personnel to properly interpret results. The LA assay is sensitive and rapid but lacks speciffcity. However, the Coccidioides Antibody EIA is a sensitive, speciffc, and rapid test for the qualitative detection of IgM (TP antigen) and IgG (CF antigen) antibodies from Coccidioides.

Citations

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Background

Coccidioides is a genus of pathogenic fungi that cause a respiratory infection known as coccidioidomycosis, or Valley fever. The two primary species within this genus are Coccidioides immitis and Coccidioides posadasii. These fungi are endemic to certain arid regions, primarily in the southwestern United States, Mexico, and parts of Central and South America. Coccidioides infections have also been reported in other regions through travel-related cases.

The life cycle of Coccidioides begins as mycelia, which are filamentous structures, in the environment and laboratory. These mycelia grow through apical extension and form septae. After a period of time, typically a week, mycelial cells undergo autolysis, resulting in the formation of barrel-shaped arthroconidia. These arthroconidia are easily aerosolized and can be inhaled by humans and other mammals. Once the arthroconidia reach the terminal bronchioles of the lungs, they transform into spherules. Spherules are spherical structures that grow to a diameter of 75 microns and internally divide into compartments containing endospores. The rupture of spherules releases endospores into the alveolar sacs, triggering an acute inflammatory response. The immune system, particularly T-helper 2 cells, plays a crucial role in combating Coccidioides infections.

saprobic life cycle and parasitic life cycle of Coccidioides. (Donovan F M, 2019) Figure 1. Life cycle of Coccidioides.
(Source: Donovan, F. M. et al., 2019)

Coccidioidomycosis can manifest in several clinical forms, ranging from a mild, self-limiting flu-like illness to severe pulmonary disease and disseminated infections involving other organs. Most infections are asymptomatic, but when symptoms occur, they can resemble community-acquired bacterial pneumonia. Common symptoms include fever, cough, shortness of breath, chest pain, headache, weight loss, and rash. The severity and extent of the infection depend on various factors, including the immune status of the individual and the inoculum size. Diagnosing coccidioidomycosis involves a combination of clinical evaluation, laboratory tests, and imaging studies. Laboratory tests may include serologic tests, such as enzyme immunoassays and complement fixation tests, as well as fungal culture and molecular methods for detecting Coccidioides DNA.

Alternative Names

Coccidioides Ab ELISA
Coccidioides Antibody ELISA
Cocci Antibody ELISA
Coccidioides Ab ELISA Kit
Coccidioides Antibody ELISA Kit
Cocci Antibody ELISA Kit

References

1. Donovan F M, et al. Early events in coccidioidomycosis. Clinical Microbiology Reviews. 2019, 33(1): 10.1128/cmr. 00112-19.

Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Virulence Mechanisms of Coccidioides

Annals of the New York Academy of Sciences

Authors: HUNG C Y U, Xue J, Cole G T.

Abstract

Coccidioides is a fungal respiratory pathogen of humans that can cause disease in both immunosuppressed and immunocompetent individuals. We describe here three mechanisms by which the pathogen survives in the hostile host environment: production of a dominant spherule outer wall glycoprotein (SOWgp) that modulates host immune response and results in compromised cell-mediated immunity to coccidioidal infection, depletion of SOWgp presentation on the surface of endospores, which prevents host recognition of the pathogen when the fungal cells are most vulnerable to phagocytic defenses, and induction of elevated production of host arginase I and coccidioidal urease, which contribute to tissue damage at sites of infection. Arginase I competes with inducible nitric oxide synthase (iNOS) in macrophages for the common substrate, L-arginine, and thereby reduces nitric oxide (NO) production and increases the synthesis of host orinithine and urea. Host-derived L-ornithine may promote pathogen growth and proliferation by providing a pool of the monoamine, which could be taken up and used for synthesis of polyamines via metabolic pathways of the parasitic cells. We have shown that high concentrations of Coccidioides- and host-derived urea at infection sites in the presence of urease produced and released by the pathogen, results in secretion of ammonia and contributes to alkalinization of the microenvironment. We propose that ammonia and enzymatically active urease released from spherules during the parasitic cycle of Coccidioides exacerbate the severity of coccidioidal infection by contributing to a compromised immune response to infection and damage of host tissue at foci of infection.

Coccidioides Species: A Review of Basic Research: 2022

Journal of Fungi

Authors: Kirkland, T. N., Stevens, D. A., Hung, C. Y., Beyhan, S., Taylor, J. W., Shubitz, L. F., Fierer, J.

Abstract

Coccidioides immitis and posadasii are closely related fungal species that cause coccidioidomycosis. These dimorphic organisms cause disease in immunocompetent as well as immunocompromised individuals and as much as 40% of the population is infected in the endemic area. Although most infections resolve spontaneously, the infection can be prolonged and, in some instances, fatal. Coccidioides has been studied for more than 100 years and many aspects of the organism and the disease it causes have been investigated. There are over 500 manuscripts concerning Coccidioides (excluding clinical articles) referenced in PubMed over the past 50 years, so there is a large body of evidence to review. We reviewed the most accurate and informative basic research studies of these fungi including some seminal older studies as well as an extensive review of current research. This is an attempt to gather the most important basic research studies about this fungus into one publication. To focus this review, we will discuss the mycology of the organism exclusively rather than the studies of the host response or clinical studies. We hope that this review will be a useful resource to those interested in Coccidioides and coccidioidomycosis.