Key result
A combined immunomagnetic separation-molecular beacon-real-time RT-PCR assay successfully detected as few as 1 PFU of Hepatitis A virus RNA and 20 PFU in seeded groundwater samples.
A novel molecular-beacon-based real-time RT-PCR assay combined with immunomagnetic separation allows for rapid and sensitive detection of Hepatitis A virus in environmental water samples.
May aid environmental HAV surveillance; leaves open real-world validation and clinical adoption.
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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Galil et al. (2004) studied Hepatitis A Virus detection. Combined IMS-MB-RT-PCR assay was evaluated on Detection limit of HAV. A combined immunomagnetic separation-molecular beacon-real-time RT-PCR assay successfully detected as few as 1 PFU of Hepatitis A virus RNA and 20 PFU in seeded groundwater samples.
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