PCR and blot hybridization for rapid identification of Haloferax species.
Based on the amplification of a 16S rDNA, a PCR assay for the identification of species of Haloferax to genus level was performed. Two variable regions of the 16S rDNA in Haloferax spp. were selected as genus-specific primers for the PCR assay and hybridization probe. Five genera of halophilic Archaea and Escherichia coli were examined as outside groups. Using this approach, all strains of Haloferax spp. were positive. In contrast, all species belonging to the most closely related genera, including Natrinema, Halorubrum, Halobacterium, and Haloarcula, were negative. In addition, the mass bloom of halophilic Archaea that develops in the El-Mallahet saltern of Alexandria City was positive using the same approach. This assay, which does not require pure cultures of microorganisms, is a specific and rapid method for identifying Haloferax spp. in hypersaline environments.
Asker
Dalal
D
Food Science and Technology, Faculty of Agriculture, Alexandria University, Egypt.
Ohta
Yoshiyuki
Y
D019641:DNA, Archaeal / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D019621:Haloferax / Q000145:classification* / Q000235:genetics; D015151:Immunoblotting / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013997:Time Factors
D019641:DNA, Archaeal; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S