Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal Samples
Human intestinal schistosomiasis is caused by the blood fluke, Schistosoma mansoni. With intensified efforts to control schistosomiasis by mass drug administration using praziquantel (PZQ), there is an urgent need to have accessible, quality-assured diagnostic tests for case detection and disease su...
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| Format: | Article |
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Wiley
2018-01-01
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| Series: | Journal of Parasitology Research |
| Online Access: | http://dx.doi.org/10.1155/2018/1267826 |
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| author | Ibrahim N. Mwangi Eric L. Agola Robert M. Mugambi Esther A. Shiraho Gerald M. Mkoji |
| author_facet | Ibrahim N. Mwangi Eric L. Agola Robert M. Mugambi Esther A. Shiraho Gerald M. Mkoji |
| author_sort | Ibrahim N. Mwangi |
| collection | DOAJ |
| description | Human intestinal schistosomiasis is caused by the blood fluke, Schistosoma mansoni. With intensified efforts to control schistosomiasis by mass drug administration using praziquantel (PZQ), there is an urgent need to have accessible, quality-assured diagnostic tests for case detection and disease surveillance and for monitoring efficacy of treatment and other interventions. Current diagnostic tools are limited by suboptimal sensitivity, slow turn-around-time, affordability, and inability to distinguish current from past infections. We describe a simple and rapid diagnostic assay, based on the loop-mediated isothermal amplification (LAMP) technology for diagnosis of S. mansoni infection in human faecal samples. The LAMP primers used in this assay were previously described and they target a 121-bp DNA repeat sequence in S. mansoni. The LAMP assay was optimized at an isothermal temperature of 63°C for 1 hour. The amplified DNA was either visualized under ultraviolet light after electrophoresis or by directly observing the color change after staining the amplicons with CYBR Green dye. The LAMP assay was evaluated against the microscopy-based procedure and the results were analysed using Cohen’s kappa coefficient to determine the degree of agreement between the two techniques. The LAMP assay reliably detected S. mansoni ova DNA in faecal samples and parasite DNA in amounts as low as 32fg. When the assay was tested for specificity against other faecal-based soil-transmitted helminths (STH), no cross-reactivity was observed. The LAMP assay was superior to the Kato-Katz assay with a 97% specificity; a high positivity score reliably detecting S. mansoni and a Kappa Coefficient of 0.9 suggested an exceptional agreement between the two techniques. The LAMP assay developed has great potential for application in field settings to support S. mansoni control and elimination campaigns. |
| format | Article |
| id | doaj-art-0f62809e72254604a196030ff0fb75e5 |
| institution | Kabale University |
| issn | 2090-0023 2090-0031 |
| language | English |
| publishDate | 2018-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Parasitology Research |
| spelling | doaj-art-0f62809e72254604a196030ff0fb75e52025-02-03T01:11:56ZengWileyJournal of Parasitology Research2090-00232090-00312018-01-01201810.1155/2018/12678261267826Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal SamplesIbrahim N. Mwangi0Eric L. Agola1Robert M. Mugambi2Esther A. Shiraho3Gerald M. Mkoji4Center for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840, 00200 Nairobi, KenyaCenter for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840, 00200 Nairobi, KenyaInstitute of Tropical Medicine and Infectious Diseases, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000, 00200 Nairobi, KenyaInstitute of Tropical Medicine and Infectious Diseases, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000, 00200 Nairobi, KenyaCenter for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840, 00200 Nairobi, KenyaHuman intestinal schistosomiasis is caused by the blood fluke, Schistosoma mansoni. With intensified efforts to control schistosomiasis by mass drug administration using praziquantel (PZQ), there is an urgent need to have accessible, quality-assured diagnostic tests for case detection and disease surveillance and for monitoring efficacy of treatment and other interventions. Current diagnostic tools are limited by suboptimal sensitivity, slow turn-around-time, affordability, and inability to distinguish current from past infections. We describe a simple and rapid diagnostic assay, based on the loop-mediated isothermal amplification (LAMP) technology for diagnosis of S. mansoni infection in human faecal samples. The LAMP primers used in this assay were previously described and they target a 121-bp DNA repeat sequence in S. mansoni. The LAMP assay was optimized at an isothermal temperature of 63°C for 1 hour. The amplified DNA was either visualized under ultraviolet light after electrophoresis or by directly observing the color change after staining the amplicons with CYBR Green dye. The LAMP assay was evaluated against the microscopy-based procedure and the results were analysed using Cohen’s kappa coefficient to determine the degree of agreement between the two techniques. The LAMP assay reliably detected S. mansoni ova DNA in faecal samples and parasite DNA in amounts as low as 32fg. When the assay was tested for specificity against other faecal-based soil-transmitted helminths (STH), no cross-reactivity was observed. The LAMP assay was superior to the Kato-Katz assay with a 97% specificity; a high positivity score reliably detecting S. mansoni and a Kappa Coefficient of 0.9 suggested an exceptional agreement between the two techniques. The LAMP assay developed has great potential for application in field settings to support S. mansoni control and elimination campaigns.http://dx.doi.org/10.1155/2018/1267826 |
| spellingShingle | Ibrahim N. Mwangi Eric L. Agola Robert M. Mugambi Esther A. Shiraho Gerald M. Mkoji Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal Samples Journal of Parasitology Research |
| title | Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal Samples |
| title_full | Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal Samples |
| title_fullStr | Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal Samples |
| title_full_unstemmed | Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal Samples |
| title_short | Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Diagnosis of Schistosoma mansoni Infection in Faecal Samples |
| title_sort | development and evaluation of a loop mediated isothermal amplification assay for diagnosis of schistosoma mansoni infection in faecal samples |
| url | http://dx.doi.org/10.1155/2018/1267826 |
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