Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept Study
The measles virus is highly contagious, and efforts to simplify its diagnosis are essential. A reverse transcriptase/recombinase polymerase amplification assay coupled with CRISPR/Cas12a and an immunochromatographic lateral flow detection (RT-RPA-CRISPR-LFD) was developed for the simple visual detec...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/14/5/517 |
| _version_ | 1797264678669254656 |
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| author | Elena Pinchon Steven Henry Fanny Leon Chantal Fournier-Wirth Vincent Foulongne Jean-François Cantaloube |
| author_facet | Elena Pinchon Steven Henry Fanny Leon Chantal Fournier-Wirth Vincent Foulongne Jean-François Cantaloube |
| author_sort | Elena Pinchon |
| collection | DOAJ |
| description | The measles virus is highly contagious, and efforts to simplify its diagnosis are essential. A reverse transcriptase/recombinase polymerase amplification assay coupled with CRISPR/Cas12a and an immunochromatographic lateral flow detection (RT-RPA-CRISPR-LFD) was developed for the simple visual detection of measles virus. The assay was performed in less than 1 h at an optimal temperature of 42 °C. The detection limit of the assay was 31 copies of an RNA standard in the reaction tube. The diagnostic performances were evaluated on a panel of 27 measles virus RT-PCR-positive samples alongside 29 measles virus negative saliva samples. The sensitivity and specificity were 96% (95% CI, 81–99%) and 100% (95% CI, 88–100%), respectively, corresponding to an accuracy of 98% (95% CI, 94–100%; <i>p</i> < 0.0001). This method will open new perspectives in the development of the point-of-care testing diagnosis of measles. |
| first_indexed | 2024-04-25T00:32:43Z |
| format | Article |
| id | doaj.art-e5e747eb5eec4eb4bded0420060b8648 |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-04-25T00:32:43Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-e5e747eb5eec4eb4bded0420060b86482024-03-12T16:42:02ZengMDPI AGDiagnostics2075-44182024-02-0114551710.3390/diagnostics14050517Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept StudyElena Pinchon0Steven Henry1Fanny Leon2Chantal Fournier-Wirth3Vincent Foulongne4Jean-François Cantaloube5Pathogénèse et Contrôle des Infections Chroniques et Emergentes, Etablissement Français du Sang, Université de Montpellier, Inserm, 34184 Montpellier, FrancePathogénèse et Contrôle des Infections Chroniques et Emergentes, Etablissement Français du Sang, Université de Montpellier, Inserm, 34184 Montpellier, FrancePathogénèse et Contrôle des Infections Chroniques et Emergentes, Etablissement Français du Sang, Université de Montpellier, Inserm, 34184 Montpellier, FrancePathogénèse et Contrôle des Infections Chroniques et Emergentes, Etablissement Français du Sang, Université de Montpellier, Inserm, 34184 Montpellier, FrancePathogénèse et Contrôle des Infections Chroniques et Emergentes, Etablissement Français du Sang, Université de Montpellier, Inserm, 34184 Montpellier, FrancePathogénèse et Contrôle des Infections Chroniques et Emergentes, Etablissement Français du Sang, Université de Montpellier, Inserm, 34184 Montpellier, FranceThe measles virus is highly contagious, and efforts to simplify its diagnosis are essential. A reverse transcriptase/recombinase polymerase amplification assay coupled with CRISPR/Cas12a and an immunochromatographic lateral flow detection (RT-RPA-CRISPR-LFD) was developed for the simple visual detection of measles virus. The assay was performed in less than 1 h at an optimal temperature of 42 °C. The detection limit of the assay was 31 copies of an RNA standard in the reaction tube. The diagnostic performances were evaluated on a panel of 27 measles virus RT-PCR-positive samples alongside 29 measles virus negative saliva samples. The sensitivity and specificity were 96% (95% CI, 81–99%) and 100% (95% CI, 88–100%), respectively, corresponding to an accuracy of 98% (95% CI, 94–100%; <i>p</i> < 0.0001). This method will open new perspectives in the development of the point-of-care testing diagnosis of measles.https://www.mdpi.com/2075-4418/14/5/517measles virusreverse transcriptionrecombinase polymerase amplificationCRISPR/Cas12 |
| spellingShingle | Elena Pinchon Steven Henry Fanny Leon Chantal Fournier-Wirth Vincent Foulongne Jean-François Cantaloube Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept Study Diagnostics measles virus reverse transcription recombinase polymerase amplification CRISPR/Cas12 |
| title | Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept Study |
| title_full | Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept Study |
| title_fullStr | Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept Study |
| title_full_unstemmed | Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept Study |
| title_short | Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with CRISPR/Cas12a and a Lateral Flow Detection: A Proof-of-Concept Study |
| title_sort | rapid detection of measles virus using reverse transcriptase recombinase polymerase amplification coupled with crispr cas12a and a lateral flow detection a proof of concept study |
| topic | measles virus reverse transcription recombinase polymerase amplification CRISPR/Cas12 |
| url | https://www.mdpi.com/2075-4418/14/5/517 |
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