A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease Virus
Visual loop-mediated isothermal amplification (LAMP) is qualified to be applied in the field to detect pathogens due to its simplicity, rapidity and cost saving. However, the color changes in currently reported visual reverse transcription LAMP (RT-LAMP) for foot-and-mouth disease virus (FMDV) detec...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-09-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/14/9/1982 |
| _version_ | 1797481377015267328 |
|---|---|
| author | Jie Zhang Qian Hou Weimin Ma Danian Chen Weibing Zhang Ashenafi Kiros Wubshet Yaozhong Ding Miaomiao Li Qian Li Jiao Chen Junfei Dai Guohua Wu Ziteng Zhang Alexei D. Zaberezhny Zygmunt Pejsak Kazimierz Tarasiuk Muhammad Umar Zafar Khan Yang Wang Jijun He Yongsheng Liu |
| author_facet | Jie Zhang Qian Hou Weimin Ma Danian Chen Weibing Zhang Ashenafi Kiros Wubshet Yaozhong Ding Miaomiao Li Qian Li Jiao Chen Junfei Dai Guohua Wu Ziteng Zhang Alexei D. Zaberezhny Zygmunt Pejsak Kazimierz Tarasiuk Muhammad Umar Zafar Khan Yang Wang Jijun He Yongsheng Liu |
| author_sort | Jie Zhang |
| collection | DOAJ |
| description | Visual loop-mediated isothermal amplification (LAMP) is qualified to be applied in the field to detect pathogens due to its simplicity, rapidity and cost saving. However, the color changes in currently reported visual reverse transcription LAMP (RT-LAMP) for foot-and-mouth disease virus (FMDV) detection are not so obvious to the naked eye, so interpretation of results is troublesome. In this study, a new naked-eye visual RT-LAMP to detect all seven distinct serotypes of FMDV was established based on the 3D genes by using pH-sensitive neutral red as the indicator, rendering a sharp contrast of color changes between the negative (light orange) and the positive (pink). Analytical sensitivity tests showed that the detection limit of the visual RT-LAMP was 10<sup>4</sup> copies/µL while those were 10<sup>3</sup> and 10<sup>4</sup> copies/µL for the RT-qPCR and conventional RT-PCR methods, respectively. Specificity tests proved that the established visual RT-LAMP assay had no cross-reactivity with other common livestock viruses. Furthermore, the analysis of 59 clinical samples showed 98.31% and 100% concordance with the RT-qPCR and the RT-PCR, respectively. The pan-serotypic FMD visual RT-LAMP assay could be suitable for a pen-side test of all seven serotypes of FMDV because the results could be easily distinguished by the naked eye without the requirement of complicated instruments and professional technicians. Hence, the novel method may have a promising prospect in field tests which exert an important role in monitoring, preventing, and controlling FMD, especially in regions with no PCR or qPCR instrument available. |
| first_indexed | 2024-03-09T22:13:41Z |
| format | Article |
| id | doaj.art-c6d6a89f4fd04b5b8e0ba73cdaa736a6 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T22:13:41Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-c6d6a89f4fd04b5b8e0ba73cdaa736a62023-11-23T19:27:33ZengMDPI AGViruses1999-49152022-09-01149198210.3390/v14091982A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease VirusJie Zhang0Qian Hou1Weimin Ma2Danian Chen3Weibing Zhang4Ashenafi Kiros Wubshet5Yaozhong Ding6Miaomiao Li7Qian Li8Jiao Chen9Junfei Dai10Guohua Wu11Ziteng Zhang12Alexei D. Zaberezhny13Zygmunt Pejsak14Kazimierz Tarasiuk15Muhammad Umar Zafar Khan16Yang Wang17Jijun He18Yongsheng Liu19Hebei Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066004, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaHebei Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066004, ChinaFederal State Budgetary Institution, All-Russian Research and Technological Institute of Biological Industry (VNITIBP), Moscow 141142, RussiaDepartment of Infectious and Parasitic Diseases, University Center of Veterinary Medicine Jagiellonian University—Agriculture Universities, 31-120 Krakow, PolandDepartment of Infectious and Parasitic Diseases, University Center of Veterinary Medicine Jagiellonian University—Agriculture Universities, 31-120 Krakow, PolandInstitute of Microbiology, University of Agriculture Faisalabad, Punjab 38040, PakistanState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaState Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaHebei Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066004, ChinaVisual loop-mediated isothermal amplification (LAMP) is qualified to be applied in the field to detect pathogens due to its simplicity, rapidity and cost saving. However, the color changes in currently reported visual reverse transcription LAMP (RT-LAMP) for foot-and-mouth disease virus (FMDV) detection are not so obvious to the naked eye, so interpretation of results is troublesome. In this study, a new naked-eye visual RT-LAMP to detect all seven distinct serotypes of FMDV was established based on the 3D genes by using pH-sensitive neutral red as the indicator, rendering a sharp contrast of color changes between the negative (light orange) and the positive (pink). Analytical sensitivity tests showed that the detection limit of the visual RT-LAMP was 10<sup>4</sup> copies/µL while those were 10<sup>3</sup> and 10<sup>4</sup> copies/µL for the RT-qPCR and conventional RT-PCR methods, respectively. Specificity tests proved that the established visual RT-LAMP assay had no cross-reactivity with other common livestock viruses. Furthermore, the analysis of 59 clinical samples showed 98.31% and 100% concordance with the RT-qPCR and the RT-PCR, respectively. The pan-serotypic FMD visual RT-LAMP assay could be suitable for a pen-side test of all seven serotypes of FMDV because the results could be easily distinguished by the naked eye without the requirement of complicated instruments and professional technicians. Hence, the novel method may have a promising prospect in field tests which exert an important role in monitoring, preventing, and controlling FMD, especially in regions with no PCR or qPCR instrument available.https://www.mdpi.com/1999-4915/14/9/1982foot-and-mouth disease virusloop-mediated isothermal amplificationnaked-eye visualizationpen-side test3D gene |
| spellingShingle | Jie Zhang Qian Hou Weimin Ma Danian Chen Weibing Zhang Ashenafi Kiros Wubshet Yaozhong Ding Miaomiao Li Qian Li Jiao Chen Junfei Dai Guohua Wu Ziteng Zhang Alexei D. Zaberezhny Zygmunt Pejsak Kazimierz Tarasiuk Muhammad Umar Zafar Khan Yang Wang Jijun He Yongsheng Liu A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease Virus Viruses foot-and-mouth disease virus loop-mediated isothermal amplification naked-eye visualization pen-side test 3D gene |
| title | A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease Virus |
| title_full | A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease Virus |
| title_fullStr | A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease Virus |
| title_full_unstemmed | A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease Virus |
| title_short | A Naked-Eye Visual Reverse Transcription Loop-Mediated Isothermal Amplification with Sharp Color Changes for Potential Pen-Side Test of Foot-and-Mouth Disease Virus |
| title_sort | naked eye visual reverse transcription loop mediated isothermal amplification with sharp color changes for potential pen side test of foot and mouth disease virus |
| topic | foot-and-mouth disease virus loop-mediated isothermal amplification naked-eye visualization pen-side test 3D gene |
| url | https://www.mdpi.com/1999-4915/14/9/1982 |
| work_keys_str_mv | AT jiezhang anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT qianhou anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT weiminma anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT danianchen anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT weibingzhang anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT ashenafikiroswubshet anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT yaozhongding anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT miaomiaoli anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT qianli anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT jiaochen anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT junfeidai anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT guohuawu anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT zitengzhang anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT alexeidzaberezhny anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT zygmuntpejsak anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT kazimierztarasiuk anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT muhammadumarzafarkhan anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT yangwang anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT jijunhe anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT yongshengliu anakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT jiezhang nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT qianhou nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT weiminma nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT danianchen nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT weibingzhang nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT ashenafikiroswubshet nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT yaozhongding nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT miaomiaoli nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT qianli nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT jiaochen nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT junfeidai nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT guohuawu nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT zitengzhang nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT alexeidzaberezhny nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT zygmuntpejsak nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT kazimierztarasiuk nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT muhammadumarzafarkhan nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT yangwang nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT jijunhe nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus AT yongshengliu nakedeyevisualreversetranscriptionloopmediatedisothermalamplificationwithsharpcolorchangesforpotentialpensidetestoffootandmouthdiseasevirus |