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Journal of Parasitology Research - Volume 2016 2016, Article ID 7376207, 7 pages -

Research Article

Institute of Tropical Medicine and Infectious Diseases, Jomo Kenyatta University of Agriculture and Technology, P.O. Box 62000-00200, Nairobi, Kenya

Centre for Biotechnology Research and Development, Kenya Medical Research Institute KEMRI, P.O. Box 54840-00200 Nairobi, Kenya

Vector Borne Disease Control Unit Laboratory, Msambweni County Referral Hospital, P.O. Box 8, Msambweni, Kenya

Received 3 May 2016; Revised 26 August 2016; Accepted 18 September 2016

Academic Editor: Tarig B. Higazi

Copyright © 2016 Esther A. Shiraho et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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.





Autor: Esther A. Shiraho, Agola L. Eric, Ibrahim N. Mwangi, Geoffrey M. Maina, Joseph M. Kinuthia, Martin W. Mutuku, Robert M. Muga

Fuente: https://www.hindawi.com/



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