Publication: Loop-mediated Isothermal Amplification (LAMP) for Identification of Pythium insidiosum
Issued Date
2020-12-01
Resource Type
ISSN
18783511
12019712
12019712
Other identifier(s)
2-s2.0-85094590497
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
International Journal of Infectious Diseases. Vol.101, (2020), 149-159
Suggested Citation
Zin Mar Htun, Tiwa Rotchanapreeda, Thidarat Rujirawat, Tassanee Lohnoo, Wanta Yingyong, Yothin Kumsang, Pattarana Sae-Chew, Penpan Payattikul, Chompoonek Yurayart, Orawan Limsivilai, Piengchan Sonthayanon, Suthee Mangmee, Piriyaporn Chongtrakool, Theerapong Krajaejun Loop-mediated Isothermal Amplification (LAMP) for Identification of Pythium insidiosum. International Journal of Infectious Diseases. Vol.101, (2020), 149-159. doi:10.1016/j.ijid.2020.09.1430 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/60020
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Loop-mediated Isothermal Amplification (LAMP) for Identification of Pythium insidiosum
Abstract
© 2020 The Author(s) Objective: Pythium insidiosum causes a life-threatening condition called pythiosis. High morbidity and mortality of pythiosis are consequences of delayed diagnosis. We aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the rapid detection of P. insidiosum for use in remote areas, where pythiosis is prevalent. Methods: We designed four LAMP primers to amplify the rDNA sequence. A side-by-side comparison evaluated performances of LAMP and the previously-established multiplex PCR (M-PCR), using gDNA samples extracted from colonies of P. insidiosum (n = 28) and other fungi (n = 54), and tissues of animals with (n = 16) or without (n = 13) pythiosis. Results: LAMP demonstrated a 50% shorter assay duration (1.5 h) and a 10-fold lower limit of detection (10-4 ng) than did M-PCR. Based on colony-extracted gDNAs, LAMP and M-PCR correctly reported P. insidiosum in all 28 samples, providing 100% sensitivity. While M-PCR did not amplify all fungal controls (100% specificity), LAMP falsely detected one organism (98% specificity). Based on the clinical samples, LAMP and M-PCR provided an equivalently-high specificity (100%). However, LAMP showed a markedly-higher sensitivity than that of M-PCR (88% vs. 56%). Conclusions: LAMP is a simple, useful, efficient assay for the detection of P. insidiosum in clinical specimens and pure cultures in resource-limited laboratories.