Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis
Abstract 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 employin...
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Nature Portfolio
2025-01-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-025-86691-8 |
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| author | Nuttapon Jirakittiwut Panuwat Sathianpitayakul Pitak Santanirand Yukihiro Akeda Tirayut Vilaivan Panan Ratthawongjirakul |
| author_facet | Nuttapon Jirakittiwut Panuwat Sathianpitayakul Pitak Santanirand Yukihiro Akeda Tirayut Vilaivan Panan Ratthawongjirakul |
| author_sort | Nuttapon Jirakittiwut |
| collection | DOAJ |
| description | Abstract 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. |
| format | Article |
| id | doaj-art-2d14f5ddc0bd4b318a3031ec54f99f74 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-2d14f5ddc0bd4b318a3031ec54f99f742025-01-26T12:33:10ZengNature PortfolioScientific Reports2045-23222025-01-0115111110.1038/s41598-025-86691-8Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosisNuttapon Jirakittiwut0Panuwat Sathianpitayakul1Pitak Santanirand2Yukihiro Akeda3Tirayut Vilaivan4Panan Ratthawongjirakul5Faculty of Allied Health Sciences, Center of Excellence for Innovative Diagnosis of Antimicrobial Resistance, Chulalongkorn UniversityMicrobiology Unit, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityMicrobiology Unit, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityDepartment of Bacteriology I, National Institute of Infectious Diseases (NIID)Organic Synthesis Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn UniversityFaculty of Allied Health Sciences, Center of Excellence for Innovative Diagnosis of Antimicrobial Resistance, Chulalongkorn UniversityAbstract 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.https://doi.org/10.1038/s41598-025-86691-8Peptide nucleic acidRecombinase polymerase amplificationRifampicin-resistant tuberculosisPaper-based sensor |
| spellingShingle | Nuttapon Jirakittiwut Panuwat Sathianpitayakul Pitak Santanirand Yukihiro Akeda Tirayut Vilaivan Panan Ratthawongjirakul Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis Scientific Reports Peptide nucleic acid Recombinase polymerase amplification Rifampicin-resistant tuberculosis Paper-based sensor |
| title | Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis |
| title_full | Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis |
| title_fullStr | Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis |
| title_full_unstemmed | Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis |
| title_short | Peptide nucleic acid-immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin-resistant tuberculosis |
| title_sort | peptide nucleic acid immobilised paper combined with multiplex recombinase polymerase amplification for the ultrasensitive and rapid detection of rifampicin resistant tuberculosis |
| topic | Peptide nucleic acid Recombinase polymerase amplification Rifampicin-resistant tuberculosis Paper-based sensor |
| url | https://doi.org/10.1038/s41598-025-86691-8 |
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