Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples
Ascaris lumbricoides is a nematode parasite that causes the common tropical infection ascariasis in humans. It is also considered among the neglected tropical diseases. Diagnosis relies mainly on microscopy-based methods which are laborious, are limited by low sensitivity, and require high expertise...
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doaj-dee47c47b73b4b36a8565052e99266612020-11-25T00:17:13ZengHindawi LimitedJournal of Parasitology Research2090-00232090-00312016-01-01201610.1155/2016/73762077376207Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal SamplesEsther A. Shiraho0Agola L. Eric1Ibrahim N. Mwangi2Geoffrey M. Maina3Joseph M. Kinuthia4Martin W. Mutuku5Robert M. Mugambi6Jackson M. Mwandi7Gerald M. Mkoji8Institute of Tropical Medicine and Infectious Diseases, Jomo Kenyatta University of Agriculture and Technology, P.O. Box 62000-00200, Nairobi, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, KenyaVector Borne Disease Control Unit Laboratory, Msambweni County Referral Hospital, P.O. Box 8, Msambweni, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, KenyaAscaris lumbricoides is a nematode parasite that causes the common tropical infection ascariasis in humans. It is also considered among the neglected tropical diseases. Diagnosis relies mainly on microscopy-based methods which are laborious, are limited by low sensitivity, and require high expertise. We have developed a loop mediated isothermal amplification (LAMP) for diagnosis of ascariasis in fecal samples, based on the first internal transcribed (ITS-1) spacer region of the ribosomal DNA. We used Primer Explorer V4 software to design primers. Ascaris adult and ova were obtained from naturally infected school children, whose parents/guardians gave consent for their participation in the study. Genomic DNA was extracted using alkaline lysis method and amplified by LAMP at 63°C for 45 minutes. LAMP products were visualized by naked eyes after adding SYBR Green dye and also on agarose gel. LAMP successfully and reliably detected Ascaris DNA from a single egg and in fecal samples. The assay specifically detected Ascaris DNA without amplifying DNA from ova of other parasites which commonly coexist with A. lumbricoides in feces. The developed LAMP assay has great potential for use in ascariasis diagnosis at the point of care and in low infection intensity situation that characterize control and elimination campaigns.http://dx.doi.org/10.1155/2016/7376207 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Esther A. Shiraho Agola L. Eric Ibrahim N. Mwangi Geoffrey M. Maina Joseph M. Kinuthia Martin W. Mutuku Robert M. Mugambi Jackson M. Mwandi Gerald M. Mkoji |
spellingShingle |
Esther A. Shiraho Agola L. Eric Ibrahim N. Mwangi Geoffrey M. Maina Joseph M. Kinuthia Martin W. Mutuku Robert M. Mugambi Jackson M. Mwandi Gerald M. Mkoji Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples Journal of Parasitology Research |
author_facet |
Esther A. Shiraho Agola L. Eric Ibrahim N. Mwangi Geoffrey M. Maina Joseph M. Kinuthia Martin W. Mutuku Robert M. Mugambi Jackson M. Mwandi Gerald M. Mkoji |
author_sort |
Esther A. Shiraho |
title |
Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples |
title_short |
Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples |
title_full |
Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples |
title_fullStr |
Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples |
title_full_unstemmed |
Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples |
title_sort |
development of a loop mediated isothermal amplification for diagnosis of ascaris lumbricoides in fecal samples |
publisher |
Hindawi Limited |
series |
Journal of Parasitology Research |
issn |
2090-0023 2090-0031 |
publishDate |
2016-01-01 |
description |
Ascaris lumbricoides is a nematode parasite that causes the common tropical infection ascariasis in humans. It is also considered among the neglected tropical diseases. Diagnosis relies mainly on microscopy-based methods which are laborious, are limited by low sensitivity, and require high expertise. We have developed a loop mediated isothermal amplification (LAMP) for diagnosis of ascariasis in fecal samples, based on the first internal transcribed (ITS-1) spacer region of the ribosomal DNA. We used Primer Explorer V4 software to design primers. Ascaris adult and ova were obtained from naturally infected school children, whose parents/guardians gave consent for their participation in the study. Genomic DNA was extracted using alkaline lysis method and amplified by LAMP at 63°C for 45 minutes. LAMP products were visualized by naked eyes after adding SYBR Green dye and also on agarose gel. LAMP successfully and reliably detected Ascaris DNA from a single egg and in fecal samples. The assay specifically detected Ascaris DNA without amplifying DNA from ova of other parasites which commonly coexist with A. lumbricoides in feces. The developed LAMP assay has great potential for use in ascariasis diagnosis at the point of care and in low infection intensity situation that characterize control and elimination campaigns. |
url |
http://dx.doi.org/10.1155/2016/7376207 |
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