Development of SCAR primers based on a repetitive DNA fingerprint for Escherichia coli detection

作者：Sangdee Aphidech^*; Natphosuk Sitakan; Srisathan Adunwit; Sangdee Kusavadee

来源：Journal of Microbiology, 2013, 51(1): 31-35.

DOI：10.1007/s12275-013-2244-4

• E. coli
• ERIC-PCR
• SCAR primer
• detection

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.

• 出版日期2013-2