title: Development of a Loop Mediated Isothermal Amplification for Diagnosis ofAscaris lumbricoidesin Fecal Samples.
abstract: Ascaris lumbricoidesis 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.Ascarisadult 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 detectedAscarisDNA from a single egg and in fecal samples. The assay specifically detectedAscarisDNA without amplifying DNA from ova of other parasites which commonly coexist withA. lumbricoidesin 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.
authors.affiliation: 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.
authors.forename: Esther A
authors.identifier: 0000-0003-2537-0588
authors.initials: EA
authors.lastname: Shiraho
authors.affiliation: Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, Kenya.
authors.forename: Agola L
authors.initials: AL
authors.lastname: Eric
authors.affiliation: Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, Kenya.
authors.forename: Ibrahim N
authors.initials: IN
authors.lastname: Mwangi
authors.affiliation: Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, Kenya.
authors.forename: Geoffrey M
authors.initials: GM
authors.lastname: Maina
authors.affiliation: Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, Kenya.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Kinuthia
authors.affiliation: Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, Kenya.
authors.forename: Martin W
authors.initials: MW
authors.lastname: Mutuku
authors.affiliation: Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, Kenya.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Mugambi
authors.affiliation: Vector Borne Disease Control Unit Laboratory, Msambweni County Referral Hospital, P.O. Box 8, Msambweni, Kenya.
authors.forename: Jackson M
authors.initials: JM
authors.lastname: Mwandi
authors.affiliation: Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), P.O. Box 54840-00200 Nairobi, Kenya.
authors.forename: Gerald M
authors.initials: GM
authors.lastname: Mkoji