title: Real-time nucleic acid sequence-based amplification assay for detection of hepatitis A virus.
abstract: A nucleic acid sequence-based amplification (NASBA) assay in combination with a molecular beacon was developed for the real-time detection and quantification of hepatitis A virus (HAV). A 202-bp, highly conserved 5' noncoding region of HAV was targeted. The sensitivity of the real-time NASBA 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, with only HAV positively identified. When combined with immunomagnetic separation, the NASBA assay successfully detected as few as 10 PFU from seeded lake water samples. Due to its isothermal nature, its speed, and its similar sensitivity compared to the real-time RT-PCR assay, this newly reported real-time NASBA method will have broad applications for the rapid detection of HAV in contaminated food or water.
authors.affiliation: Department of Microbiology, Mansoura University, Egypt.
authors.forename: Khaled H
authors.initials: KH
authors.lastname: Abd el-Galil
authors.forename: M A
authors.initials: MA
authors.lastname: el-Sokkary
authors.forename: S M
authors.initials: SM
authors.lastname: Kheira
authors.forename: Andre M
authors.initials: AM
authors.lastname: Salazar
authors.forename: Marylynn V
authors.initials: MV
authors.lastname: Yates
authors.forename: Wilfred
authors.initials: W
authors.lastname: Chen
authors.forename: Ashok
authors.initials: A
authors.lastname: Mulchandani
mesh_terms: D000818:Animals; D005618:Fresh Water / Q000821:virology; D030041:Hepatitis A virus / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D018189:Immunomagnetic Separation; D015335:Molecular Probes; D012367:RNA, Viral / Q000032:analysis; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D015335:Molecular Probes; D012367:RNA, Viral