The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis.
<h4>Background</h4>Lack of available sensitive point-of-care testing is one of the primary obstacles to the rapid diagnosis of leptospirosis. The purpose of this study was to test the performance of two point-of-care tests, a clustered regularly interspaced short palindromic repeat (CRIS...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2023-08-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011596&type=printable |
| _version_ | 1827801108375928832 |
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| author | Sirawit Jirawannaporn Umaporn Limothai Sasipha Tachaboon Janejira Dinhuzen Patcharakorn Kiatamornrak Watchadaporn Chaisuriyong Nattachai Srisawat |
| author_facet | Sirawit Jirawannaporn Umaporn Limothai Sasipha Tachaboon Janejira Dinhuzen Patcharakorn Kiatamornrak Watchadaporn Chaisuriyong Nattachai Srisawat |
| author_sort | Sirawit Jirawannaporn |
| collection | DOAJ |
| description | <h4>Background</h4>Lack of available sensitive point-of-care testing is one of the primary obstacles to the rapid diagnosis of leptospirosis. The purpose of this study was to test the performance of two point-of-care tests, a clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 12a (CRISPR/Cas12a) fluorescence-based diagnostic assay (FBDA), a Leptospira immunoglobulin M (IgM) rapid diagnostic test (RDT), and the two tests combined.<h4>Methodology/principal findings</h4>For the diagnosis of 171 clinical samples, a recombinase polymerase amplification (RPA)-CRISPR/Cas12a FBDA for whole blood and Leptospira IgM RDT (Medical Science Public Health, Thailand) for serum were used. The confirmed cases were determined by using any positive qPCR, microscopic agglutination test (MAT), and culture results. Diagnostic accuracy was assessed on the first day of enrollment and stratified by the day after symptom onset. The overall sensitivity of the Leptospira IgM RDT and RPA-CRISPR/Cas12a FBDA was 55.66% and 60.38%, respectively. When the two tests were combined, the sensitivity rose to 84.91%. The specificity of each test was 63.08% and 100%, respectively, and 63.08% when combined. The sensitivity of the Leptospira IgM RDT rose on days 4-6 after the onset of fever, while the RPA-CRISPR/Cas12a FBDA continued to decrease. When the two tests were combined, the sensitivity was over 80% at different days post-onset of fever.<h4>Conclusions/significance</h4>The combination of Leptospira IgM RDT and RPA-CRISPR/Cas12 FBDA exhibited significant sensitivity for the detection of leptospires at various days after the onset of fever, thereby reducing the likelihood of misdiagnosis. The combination of these assays may be suitable for early leptospirosis screening in situations with limited resources. |
| first_indexed | 2024-03-11T20:20:42Z |
| format | Article |
| id | doaj.art-b46fda9b0fdc441fa38604139c300381 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-03-11T20:20:42Z |
| publishDate | 2023-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-b46fda9b0fdc441fa38604139c3003812023-10-03T05:31:56ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352023-08-01178e001159610.1371/journal.pntd.0011596The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis.Sirawit JirawannapornUmaporn LimothaiSasipha TachaboonJanejira DinhuzenPatcharakorn KiatamornrakWatchadaporn ChaisuriyongNattachai Srisawat<h4>Background</h4>Lack of available sensitive point-of-care testing is one of the primary obstacles to the rapid diagnosis of leptospirosis. The purpose of this study was to test the performance of two point-of-care tests, a clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 12a (CRISPR/Cas12a) fluorescence-based diagnostic assay (FBDA), a Leptospira immunoglobulin M (IgM) rapid diagnostic test (RDT), and the two tests combined.<h4>Methodology/principal findings</h4>For the diagnosis of 171 clinical samples, a recombinase polymerase amplification (RPA)-CRISPR/Cas12a FBDA for whole blood and Leptospira IgM RDT (Medical Science Public Health, Thailand) for serum were used. The confirmed cases were determined by using any positive qPCR, microscopic agglutination test (MAT), and culture results. Diagnostic accuracy was assessed on the first day of enrollment and stratified by the day after symptom onset. The overall sensitivity of the Leptospira IgM RDT and RPA-CRISPR/Cas12a FBDA was 55.66% and 60.38%, respectively. When the two tests were combined, the sensitivity rose to 84.91%. The specificity of each test was 63.08% and 100%, respectively, and 63.08% when combined. The sensitivity of the Leptospira IgM RDT rose on days 4-6 after the onset of fever, while the RPA-CRISPR/Cas12a FBDA continued to decrease. When the two tests were combined, the sensitivity was over 80% at different days post-onset of fever.<h4>Conclusions/significance</h4>The combination of Leptospira IgM RDT and RPA-CRISPR/Cas12 FBDA exhibited significant sensitivity for the detection of leptospires at various days after the onset of fever, thereby reducing the likelihood of misdiagnosis. The combination of these assays may be suitable for early leptospirosis screening in situations with limited resources.https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011596&type=printable |
| spellingShingle | Sirawit Jirawannaporn Umaporn Limothai Sasipha Tachaboon Janejira Dinhuzen Patcharakorn Kiatamornrak Watchadaporn Chaisuriyong Nattachai Srisawat The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis. PLoS Neglected Tropical Diseases |
| title | The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis. |
| title_full | The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis. |
| title_fullStr | The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis. |
| title_full_unstemmed | The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis. |
| title_short | The combination of RPA-CRISPR/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis. |
| title_sort | combination of rpa crispr cas12a and leptospira igm rdt enhances the early detection of leptospirosis |
| url | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011596&type=printable |
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