Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, Kenya
Introduction. Accurate detection of filarial parasites in humans and vectors is essential for the implementation and evaluation of Global and National Programs to eliminate lymphatic filariasis. Immunological methods to detect infection are available; however, cross-reactivity issues have been repor...
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Wiley
2021-01-01
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| Series: | Journal of Parasitology Research |
| Online Access: | http://dx.doi.org/10.1155/2021/6650870 |
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| author | Kinyatta Nancy Wambua Lillian Mutahi Wilkinson Mugasa Claire Kamau Luna Wachira Dorcas Githae Rosemary Lusweti Japheth Ichugu Christine Waigi Emily Kagai Jim |
| author_facet | Kinyatta Nancy Wambua Lillian Mutahi Wilkinson Mugasa Claire Kamau Luna Wachira Dorcas Githae Rosemary Lusweti Japheth Ichugu Christine Waigi Emily Kagai Jim |
| author_sort | Kinyatta Nancy |
| collection | DOAJ |
| description | Introduction. Accurate detection of filarial parasites in humans and vectors is essential for the implementation and evaluation of Global and National Programs to eliminate lymphatic filariasis. Immunological methods to detect infection are available; however, cross-reactivity issues have been reported in most of them. Nucleic acid-based molecular assays offer high levels of specificity and sensitivity and can be used to detect the infections. Methods. In this study, we evaluated loop-mediated isothermal amplification (LAMP) tests to amplify Wuchereria bancrofti DNA in patients’ blood. The amplicons were tested by both pH-sensitive dyes for enhanced visual detection and agarose gel electrophoresis. A closed-tube LAMP assay was also evaluated. Cohen’s Kappa statistics was used for statistical analysis of the assays. 125 patients consented for blood sampling which were used for clinical analysis of LAMP assays with the PCR method used as the “gold standard.” Results. The sensitivity of the evaluated Wuchereria bancrofti LAMP was 92.3%, with a specificity of 97.3% and kappa statistics value of 0.84, which is in a strong agreement. Conclusion. In this study, LAMP assays coupled with fluorescence dye detection have been found to be suitable for diagnosis and monitoring of Wuchereria bancrofti infections in the Kenyan population. |
| format | Article |
| id | doaj-art-fea354523df64312a39819adf7483d61 |
| institution | Kabale University |
| issn | 2090-0023 2090-0031 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Parasitology Research |
| spelling | doaj-art-fea354523df64312a39819adf7483d612025-02-03T07:23:57ZengWileyJournal of Parasitology Research2090-00232090-00312021-01-01202110.1155/2021/66508706650870Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, KenyaKinyatta Nancy0Wambua Lillian1Mutahi Wilkinson2Mugasa Claire3Kamau Luna4Wachira Dorcas5Githae Rosemary6Lusweti Japheth7Ichugu Christine8Waigi Emily9Kagai Jim10Kenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaSchool of Biological Science, University of Nairobi, 30772-00100, Nairobi, KenyaSchool of Biological Science, University of Nairobi, 30772-00100, Nairobi, KenyaCollege of Veterinary Medicine, Animal Resources and Biosecurity, Makerere University Kampala, 7062 Kampala, UgandaKenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaKenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaKenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaKenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaKenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaKenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaKenya Medical Research Institute, Centre for Biotechnology Research and Development, P.O. Box 54840-00200, Nairobi, KenyaIntroduction. Accurate detection of filarial parasites in humans and vectors is essential for the implementation and evaluation of Global and National Programs to eliminate lymphatic filariasis. Immunological methods to detect infection are available; however, cross-reactivity issues have been reported in most of them. Nucleic acid-based molecular assays offer high levels of specificity and sensitivity and can be used to detect the infections. Methods. In this study, we evaluated loop-mediated isothermal amplification (LAMP) tests to amplify Wuchereria bancrofti DNA in patients’ blood. The amplicons were tested by both pH-sensitive dyes for enhanced visual detection and agarose gel electrophoresis. A closed-tube LAMP assay was also evaluated. Cohen’s Kappa statistics was used for statistical analysis of the assays. 125 patients consented for blood sampling which were used for clinical analysis of LAMP assays with the PCR method used as the “gold standard.” Results. The sensitivity of the evaluated Wuchereria bancrofti LAMP was 92.3%, with a specificity of 97.3% and kappa statistics value of 0.84, which is in a strong agreement. Conclusion. In this study, LAMP assays coupled with fluorescence dye detection have been found to be suitable for diagnosis and monitoring of Wuchereria bancrofti infections in the Kenyan population.http://dx.doi.org/10.1155/2021/6650870 |
| spellingShingle | Kinyatta Nancy Wambua Lillian Mutahi Wilkinson Mugasa Claire Kamau Luna Wachira Dorcas Githae Rosemary Lusweti Japheth Ichugu Christine Waigi Emily Kagai Jim Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, Kenya Journal of Parasitology Research |
| title | Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, Kenya |
| title_full | Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, Kenya |
| title_fullStr | Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, Kenya |
| title_full_unstemmed | Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, Kenya |
| title_short | Optimization of a Loop-Mediated Isothermal Amplification Assay as a Point-of-Care Tool for the Detection of Wuchereria bancrofti in Human Blood in Tana River Delta, Kenya |
| title_sort | optimization of a loop mediated isothermal amplification assay as a point of care tool for the detection of wuchereria bancrofti in human blood in tana river delta kenya |
| url | http://dx.doi.org/10.1155/2021/6650870 |
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