title: Real-time PCR for differentiation of F4 (K88) variants (F4ab, F4ac, F4ad) of enterotoxigenic Escherichiacoli from diarrhoeic piglets.
abstract: A one-step real-time PCR using one set of oligonucleotide primers and three probes was developed for differentiation of F4 (K88) variants (F4ab, F4ac, F4ad) of enterotoxigenic Escherichiacoli (ETEC) from diarrhoeic pigs. The limits of detection of F4ab, F4ac and F4ad in broth dilution were 10(6), 10(5) and 10(4)colony forming units (CFU)/mL, respectively. In faecal samples spiked with E.coli, the limits of detection of F4ab, F4ac and F4ad were 10(6), 10(6) and 10(4)CFU/g faeces, respectively, without enrichment and 10(3), 10(2) and 10(2)CFU/g faeces following enrichment. In 42 ETEC field isolates from pigs in Korea encoding the F4 gene, all were identified as the F4ac variant.
authors.affiliation: Animal Disease Diagnostic Centre, National Veterinary Research and Quarantine Service, Anyang 430-757, Republic of Korea.
authors.forename: Jae-Won
authors.initials: JW
authors.lastname: Byun
authors.forename: Byeong Yeal
authors.initials: BY
authors.lastname: Jung
authors.forename: Ha-Young
authors.initials: HY
authors.lastname: Kim
authors.forename: John M
authors.initials: JM
authors.lastname: Fairbrother
authors.forename: Wan-Kyu
authors.initials: WK
authors.lastname: Lee
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D003967:Diarrhea / Q000382:microbiology / Q000662:veterinary*; D054307:Enterotoxigenic Escherichia coli / Q000235:genetics* / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D005243:Feces / Q000382:microbiology; D036781:Fimbriae Proteins / Q000235:genetics* / Q000378:metabolism; D057230:Limit of Detection; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D056910:Republic of Korea; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000382:microbiology*
chemical_list: D000942:Antigens, Bacterial; D029968:Escherichia coli Proteins; C026888:K88 antigen, E coli; D036781:Fimbriae Proteins