Open AccessRapid and sensitive method for detection of Salmonella strains using a combination of polymerase chain reaction and reverse dot‐blot hybridization
Author(s) -
Iida Kazuo,
Abe Akio,
Matsui Hidenori,
Danbara Hirofumi,
Wakayama Sachio,
Kawahara Kazuyoshi
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06568.x
Subject(s) - salmonella , dot blot , biology , polymerase chain reaction , microbiology and biotechnology , oligonucleotide , oligomer restriction , enterobacteriaceae , hybridization probe , southern blot , 16s ribosomal rna , bacteria , bacillus subtilis , molecular probe , serotype , dna , gene , escherichia coli , biochemistry , genetics
We have developed a reverse dot‐blot hybridization assay for detection of Salmonella using Salmonella ‐specific o oligonucleotide probes designed from the base sequence of the 16S rRNA gene (rDNA). The target fragment of 16S rDNA was amplified, and labelled with biotin by the polymerase chain reaction. The amplified fragment was hybridized with the membrane‐immobilized probe and the hybridization was detected by chemiluminescence. Amplified fragments from 24 different serovars of Salmonella hybridized with the probes, whereas those of species of Enterobacteriaceae, Pseudomonas aeruginosa, Bacillus subtilis , and Staphylococcus aureus failed to hybridize. By this assay, it was possible to detect in the order of 10 4 bacteria in fish meat homogenate in 10 h.