title: Multiplex approach for screening genetic markers of microbial indicators.
abstract: Genetic markers are expected to provide better specificity in epidemiological studies and potentially serve as better indicators of waterborne pathogens. Methods used to assess genetic markers of emerging microbial indicators include pulsed field gel electrophoresis, polymerase chain reaction (PCR), and microarrays. This paper outlines a high-throughput approach to screen for such genetic markers using a set of theoretical and experimental screening tools. The theoretical screening involves evaluating genes related to the ribosomal RNA and specific functions from emerging indicator groups, followed by experimental validation with appropriate sampling schemes and high-throughput and economical screening methods, such as microarrays, real time PCR, and on-chip PCR. Analysis of a wide range of samples covering temporal variability in location, host, and waterborne disease outbreaks is essential. The proposed approach is expected to shorten the time and cost associated with searching for new genetic markers of emerging indicators by at least 10-fold.
authors.affiliation: Department of Civil and Environmental Engineering, Michigan State University, East Lansing 48824, USA.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Stedtfeld
authors.forename: Sam
authors.initials: S
authors.lastname: Baushke
authors.forename: Dieter
authors.initials: D
authors.lastname: Tourlousse
authors.forename: Benli
authors.initials: B
authors.lastname: Chai
authors.forename: James R
authors.initials: JR
authors.lastname: Cole
authors.forename: Syed A
authors.initials: SA
authors.lastname: Hashsham
mesh_terms: D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D004269:DNA, Bacterial / Q000235:genetics; D005819:Genetic Markers / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D037521:Virulence Factors / Q000235:genetics; D014871:Water Microbiology / Q000592:standards*
chemical_list: D004269:DNA, Bacterial; D005819:Genetic Markers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D037521:Virulence Factors