title: Development of SCAR primers based on a repetitive DNA fingerprint for Escherichia coli detection.
abstract: The present study aimed to use enterobacterial repetitive intergenic consensus (ERIC) fingerprints to design SCAR primers for the detection of Escherichia coli. The E. coli strains were isolated from various water sources. The primary presumptive identification of E. coli was achieved using MacConkey agar. Nineteen isolates were selected and confirmed to be E. coli strains based on seven biochemical characteristics. ERIC-PCR with ERIC 1R and ERIC 2 primers were used to generate DNA fingerprints. ERIC-PCR DNA profiles showed variant DNA profiles among the tested E. coli strains and distinguished all E. coli strains from the other tested bacterial strains. A 350 bp band that predominated in five E. coli strains was used for the development of the species-specific SCAR primers EC-F1 and EC-R1. The primers showed good specificity for E. coli, with the exception of a single false positive reaction with Sh. flexneri DMST 4423. The primers were able to detect 50 pg and 10(0) CFU/ml of genomic DNA and cells of E. coli, respectively.
authors: Department of Biology, Faculty of Science, Mahasarakham University, Mahasarakham 44150, Thailand. aphidech_sangdee@yahoo.com
authors: Aphidech
authors: A
authors: Sangdee
authors: Sitakan
authors: S
authors: Natphosuk
authors: Adunwit
authors: A
authors: Srisathan
authors: Kusavadee
authors: K
authors: Sangdee
mesh_terms: D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D005189:False Positive Reactions; D058889:Molecular Typing / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity; D014871:Water Microbiology
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial