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Rapid identification of S almonella using H ektoen enteric agar and 16s ribosomal DNA probe‐gold nanoparticle immunochromatography assay in clinical faecal specimens
Author(s) -
Yeung C.Y.,
Liu C.C.,
Tseng Y.T.,
Tsai K.C.,
Hsieh M.A.,
Chan W.T.,
Liu H.L.,
Lee H.C.,
Hou S.Y.
Publication year - 2014
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12191
Subject(s) - salmonella , gold standard (test) , microbiology and biotechnology , 16s ribosomal rna , colloidal gold , agar , polymerase chain reaction , biology , ribosomal rna , bacteria , clinical microbiology , immunoassay , agar plate , antibody , medicine , gene , biochemistry , immunology , genetics , nanoparticle , materials science , nanotechnology
A rapid identification of S almonella , one of the most common foodborne pathogens worldwide, in clinical patients can enable better rational managements and prevent further outbreaks. The traditional immunochromatography using antibody–gold nanoparticles (Ab‐Au NP s), such as the home pregnancy test, has been used for the S almonella detection. In this study, we developed a new and rapid method using DNA probe‐Au NP s for the detection of 16s ribosomal DNA of S almonella . To evaluate the proposed method in clinical specimens, we performed a clinical test by identifying 159 stool samples on H ektoen agar containing black or crystalloid colonies using the method and the VITEK 2 system for confirmation. Eighty of the isolates were correctly identified as S almonella to achieve 100% sensitivity. Seventy‐five samples were correctly identified as non‐ S almonella spp., but four were incorrectly identified as S almonella . The specificity was 94·93%. The assay time is about 30 min after the DNA purification. The time‐consuming and labour‐intense biochemical tests can be replaced. We demonstrated that this assay is a rapid, convenient and cost‐effective tool for S almonella identification of clinical faecal samples, which is worth for further promotion and clinical use. This is the first application of using 16s ribosomal DNA probe‐Au‐ NP s and immunochromatography on clinical samples. Significance and Impact of the Study This is the first application of using 16s ribosomal DNA probe–gold nanoparticles and immunochromatography method on clinical samples with sensitivity 100% and specificity 94·93%. The assay time is about 30 min after the DNA purification. We find this assay a rapid, convenient, sensitive and inexpensive tool for S almonella identification of clinical faecal samples, which is worth further promotion and clinical use and can replace the traditional time‐consuming and labour‐intense biochemical tests. The potential benefit of this approach is to develop a rapid point‐of‐care test that provides results while the patient is still at the doctors' office.