Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis

Autor: Elisa Roncal, Edgar A. Florentini, Patricia Sheen, Roberto Alcántara, Daniela E. Kirwan, Ricardo Antiparra, Emily Toscano, Mirko Zimic, Katherine Vallejos, Robert H. Gilman, Noelia Angulo
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Bacterial Diseases
Immunoconjugates
Physiology
Metabolite
Antitubercular Agents
Biochemistry
Immunologic Adjuvants
chemistry.chemical_compound
Medical Conditions
Immune Physiology
Medicine and Health Sciences
Public and Occupational Health
Enzyme-Linked Immunoassays
Bovine serum albumin
Enzyme-linked immunoassays
0303 health sciences
Immune System Proteins
Multidisciplinary
biology
Chemical Synthesis
Serum Albumin
Bovine

Vaccination and Immunization
3. Good health
Actinobacteria
Infectious Diseases
Medicine
Rabbits
Research Article
medicine.drug
endocrine system
Science
Immunology
Enzyme-Linked Immunosorbent Assay
Research and Analysis Methods
Antibodies
Mycobacterium tuberculosis
03 medical and health sciences
Inhibitory Concentration 50
Pyrazinoic acid
Drug Resistance
Bacterial

Toxicity Tests
Genetics
medicine
Animals
Tuberculosis
Chemical synthesis
Antigens
Immunoassays
IC50
030304 developmental biology
Detection limit
Chromatography
Bacteria
030306 microbiology
Organisms
Immunologic adjuvants
Biology and Life Sciences
Proteins
Correction
Pyrazinamide
Tropical Diseases
biology.organism_classification
chemistry
Polyclonal antibodies
purl.org/pe-repo/ocde/ford#3.02.07 [https]
Mutation
Immunologic Techniques
biology.protein
Preventive Medicine
Zdroj: PLoS ONE
PLoS ONE, Vol 15, Iss 11, p e0241600 (2020)
ISSN: 1932-6203
Popis: Pyrazinamide (PZA) susceptibility testing in Mycobacterium tuberculosis (Mtb) is a current area of development and PZA-resistant strains are increasingly prevalent. Previous studies have demonstrated that the detection of pyrazinoic acid (POA), the metabolite produced by the deamidation of PZA, is a good predictor for PZA resistance since a resistant strain would not convert PZA into POA at a critical required rate, whereas a susceptible strain will do, expelling POA to the extracellular environment at a certain rate, and allowing for quantification of this accumulated analyte. In order to quantify POA, an indirect competitive ELISA (icELISA) test using hyperimmune polyclonal rabbit serum against POA was developed: for this purpose, pure POA was first covalently linked to the highly immunogenic Keyhole Limpet Hemocyanine, and inoculated in rabbits. A construct made of bovine serum albumin (BSA) linked to pure POA and fixed at the bottom of wells was used as a competitor against spiked samples and liquid Mtb culture supernatants. When spiked samples (commercial POA alone) were analyzed, the half maximal inhibitory concentration (IC50) was 1.16 mg/mL, the limit of detection 200 μg/mL and the assay was specific (it did not detect PZA, IC50 > 20 mg/mL). However, culture supernatants (7H9-OADC-PANTA medium) disrupted the competition and a proper icELISA curve was not obtainable. We consider that, although we have shown that it is feasible to induce antibodies against POA, matrix effects could damage its analytical usefulness; multiple, upcoming ways to solve this obstacle are suggested.
Databáze: OpenAIRE