

@article {4263,
	title = {{Genotyping male-specific RNA coliphages by hybridization with oligonucleotide probes.}},
	journal = {Applied and Environmental Microbiology},
	volume = {61},
	number = {11},
	year = {1995},
	month = {nov},
	pages = {3960{\textendash}6},
	abstract = {<p>F-specific (F+) RNA coliphages are prevalent in sewage and other fecal wastes of humans and animals. There are four antigenically distinct serogroups of F+ RNA coliphages, and those predominating in humans (groups II and III) differ from those predominating in animals (groups I and IV). Hence, it may be possible to distinguish between human and animal wastes by serotyping F+ RNA coliphage isolates. Because serotyping is laborious and requires scarce antiserum reagents, we investigated genotyping using synthetic oligonucleotide probes as an alternative approach to distinguishing the four groups of F+ RNA coliphages. Oligoprobes I, II, III, IV, A, and B were selected to detect group I, II, III, IV, I plus II, and III plus IV phages, respectively. Methods for phage transfer from zones of lysis on a host cell lawn to candidate membrane filters and fixation of genomic nucleic acid on the membranes were optimized. The oligoprobes, which were end labeled with digoxigenin, were applied in DNA-RNA hybridization, and hybrids were observed by colorimetric, immunoenzymatic detection. Of 203 isolates of F+ RNA coliphages from environmental samples of water, wastes, and shellfish, 99.5 and 96.6{\%} could be classified into each group by serotyping and genotyping, respectively. Probes A and B correctly identified 100{\%} of the isolates. On the basis of these results, this method for genotyping F+ RNA coliphages appears to be practical and reliable for typing isolates in field samples.</p>
},
	keywords = {Animals, Base Sequence, Coliphages, Coliphages: classification, Coliphages: genetics, Coliphages: isolation {\&} purification, Environmental Microbiology, Feces, Feces: virology, genotype, Humans, Molecular Sequence Data, Nucleic Acid Hybridization, Oligonucleotide Probes, Oligonucleotide Probes: genetics, RNA, Serotyping, Viral, Viral: genetics},
	issn = {0099-2240},
	url = {http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=167702{\&}tool=pmcentrez{\&}rendertype=abstract},
	author = {Hsu, F C and Shieh, Y S and van Duin, J and Beekwilder, M J and Sobsey, M D}
}
