Development of a thermostabilized, one-step, nested, tetraplex PCR assay for simultaneous identification and differentiation of Entamoeba species, Entamoeba histolytica and Entamoeba dispar from stool samples

Journal of Medical Microbiology
Phiaw Chong FooBoon Huat Lim

Abstract

Entamoeba histolytica is the only Entamoeba species that causes amoebiasis in humans. Approximately 50 million people are infected, with 100, 000 deaths annually in endemic countries. Molecular diagnosis of Entamoeba histolytica is important to differentiate it from the morphologically identical Entamoeba dispar to avoid unnecessary medication. Conventional molecular diagnostic tests require trained personnel, cold-chain transportation and/or are storage-dependent, which make them user-unfriendly. The aim of this study was to develop a thermostabilized, one-step, nested, tetraplex PCR assay for the detection of Entamoeba histolytica, Entamoeba dispar and Entamoeba species in cold-chain-free and ready-to-use form. The PCR test was designed based on the Entamoeba small subunit rRNA (SSU-rRNA) gene, which detects the presence of any Entamoeba species, and simultaneously can be used to differentiate Entamoeba histolytica from Entamoeba dispar. In addition, a pair of primers was designed to serve as an internal amplification control to help identify inhibitors in the samples. All PCR reagents together with the designed primers were thermostabilized by lyophilization and were stable at 24 °C for at least 6 months. The limit of detect...Continue Reading

References

Dec 1, 1992·International Journal for Parasitology·H TachibanaG Masuda
Mar 1, 1994·Journal of Clinical Pathology·A González-RuizD C Warhurst
Mar 1, 1997·The Journal of Infectious Diseases·R HaqueW A Petri
Nov 5, 1997·Lancet·S Hohenschild
Jun 10, 1998·Journal of Clinical Microbiology·P R KlatserA H Kolk
Jul 5, 2002·Clinical Microbiology Reviews·C Graham Clark, Louis S Diamond
Sep 23, 2003·Current Opinion in Infectious Diseases·William Stauffer, Jonathan I Ravdin
Oct 15, 2003·Clinical Microbiology Reviews·Mehmet Tanyuksel, William A Petri
Nov 26, 2004·Journal of Clinical Pathology·S J FurrowsP L Chiodini
Sep 7, 2006·Journal of Clinical Microbiology·Zulhainan HamzahPorntip Chavalitshewinkoon-Petmitr
Jul 17, 2007·Clinical Microbiology Reviews·R FotedarJ Harkness
Oct 30, 2007·Diagnostic Microbiology and Infectious Disease·Ismail AziahAsma Ismail
Dec 25, 2010·Journal of Medical Microbiology·Ang Lim ChuaPattabhiraman Lalitha

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Citations

May 20, 2014·Travel Medicine and Infectious Disease·Pak-On LeungKow-Tong Chen
Feb 15, 2015·Experimental Parasitology·Gustavo A FontechaByron Leiva
Nov 17, 2017·Clinical Microbiology Reviews·Lynne S GarciaGovinda Visvesvara
May 1, 2021·Diagnostics·Nik Mohd Noor Nik ZurainaSiti Suraiya

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