Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis.

Publication date: Jan 21, 2025

Rifampicin-resistant tuberculosis (RR-TB) is a critical issue with significant implications for patient care, public health, and TB control efforts that necessitate comprehensive strategies for detection. This study presents a novel point-of-care diagnostic tool for RR-TB detection employing a peptide nucleic acid (PNA)-paper-based sensor combined with isothermal recombinase polymerase amplification (RPA). The sensor targets mutations in codons 516, 526, and 531 of the rpoB gene, the top three common mutations associated with rifampicin-resistant strains. PNA probes specifically recognised wild-type sequences, generating a visual signal through a reverse hybridisation assay. The absence of a signal was observed when the mutant strains were detected because of the inability to bind the mutant sequence. Our proof-of-concept assay displayed high accuracy (100% for detecting mutations at codons 516, 526, and 531), a short turnaround time (110 min), no cross-reactivity with other bacterial pathogens, and ultrasensitivity. This PNA-paper-based sensor model can be a valuable diagnostic tool for RR-TB detection, providing an accessible diagnostic platform that can be advantageous in resource-limited settings where sophisticated laboratory infrastructure may be lacking.

Open Access PDF

Concepts Keywords
Isothermal Bacterial Proteins
Laboratory Bacterial Proteins
Mutant DNA-Directed RNA Polymerases
Recombinase DNA-Directed RNA Polymerases
Tuberculosis Drug Resistance, Bacterial
Humans
Mutation
Mycobacterium tuberculosis
Paper
Paper-based sensor
Peptide nucleic acid
Peptide Nucleic Acids
Peptide Nucleic Acids
Recombinase polymerase amplification
Recombinases
Recombinases
Rifampicin-resistant tuberculosis
Rifampin
Rifampin
Sensitivity and Specificity
Tuberculosis, Multidrug-Resistant

Semantics

Type Source Name
disease IDO nucleic acid
drug DRUGBANK Rifampicin
disease MESH tuberculosis
pathway KEGG Tuberculosis
disease IDO assay
drug DRUGBANK Isoniazid
disease MESH point mutations
disease MESH Infectious Diseases
drug DRUGBANK Coenzyme M
drug DRUGBANK Carboxymethylcellulose
drug DRUGBANK Water
drug DRUGBANK Medical air
drug DRUGBANK L-Lysine
drug DRUGBANK Sodium carbonate
drug DRUGBANK Phosphate ion
disease IDO colony
drug DRUGBANK Biotin
drug DRUGBANK Carbonate ion
drug DRUGBANK Alkaline Phosphatase
drug DRUGBANK Sodium Citrate
drug DRUGBANK Sodium lauryl sulfate
disease IDO drug susceptibility
disease MESH thalassemia
disease IDO process
drug DRUGBANK Isoxaflutole
drug DRUGBANK Gold
disease MESH clinical relevance
disease IDO site
drug DRUGBANK Tretamine
disease MESH Pneumonia
disease MESH pulmonary tuberculosis
disease MESH hepatitis
disease IDO cell
drug DRUGBANK Amino acids
disease IDO antibiotic resistance
pathway REACTOME Reproduction
disease MESH Tuberculosis Multidrug-Resistant

Original Article

Leave a Comment

Your email address will not be published. Required fields are marked *