Combined immunomagnetic separation-molecular beacon-reverse transcription-PCR assay for detection of hepatitis A virus from environmental samples

Applied and Environmental Microbiology
Khaled H Abd El GalilAshok Mulchandani

Abstract

In this study, a molecular-beacon-based real-time reverse transcription (RT)-PCR assay was developed to detect the presence of hepatitis A virus (HAV) in environmental samples. A 125-bp, highly conserved 5' noncoding region of HAV was targeted. The sensitivity of the real-time RT-PCR assay was tested with 10-fold dilutions of viral RNA, and a detection limit of 1 PFU was obtained. The specificity of the assay was demonstrated by testing with other environmental pathogens and indicator microorganisms, and only HAV was positively identified. When combined with immunomagnetic separation, the real-time RT-PCR assay successfully detected as few as 20 PFU in seeded groundwater samples. Because of its simplicity and specificity, this assay has broad applications for the rapid detection of HAV in contaminated foods or water.

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Citations

May 18, 2005·Journal of Virological Methods·Hilary A BrooksArun K Dhar
Apr 23, 2011·Lab on a Chip·Benjamin R SchudelPaul J A Kenis
Feb 12, 2008·Applied and Environmental Microbiology·Hsiao-Yun YehWilfred Chen
Nov 5, 2005·Applied and Environmental Microbiology·Khaled H Abd el-GalilAshok Mulchandani
Jan 19, 2006·Clinical Microbiology Reviews·Omana V NainanHarold S Margolis
Sep 24, 2008·BMC Microbiology·Véronique Apaire-MarchaisRaymond Robert
Sep 17, 2011·International Journal of Hygiene and Environmental Health·Ibrahim Ahmed HamzaMichael Wilhelm
May 12, 2009·Seminars in Cell & Developmental Biology·Hsiao-Yun YehAshok Mulchandani
Dec 13, 2006·Journal of Virological Methods·Alain HoudeYvon-Louis Trottier
Nov 19, 2011·FEMS Microbiology Reviews·David Rodríguez-LázaroWim H M van der Poel
Feb 19, 2021·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Paymaneh AtabakhshAbbas Doosti

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