Application of colorimetric microdilution plate hybridization for rapid genetic identification of 22 Mycobacterium species.
Quantitative microdilution plate hybridization was used to identify 22 Mycobacterium species. DNAs of clinical strains were rapidly extracted and labeled with photoreactive biotin. Labeled DNAs were distributed into wells of a microdilution plate in which reference DNAs had been immobilized. After 2 h of hybridization, hybridized DNAs were quantitatively detected with peroxidase-conjugated streptavidin and the substrate, tetramethylbenzidine. This method could differentiate among 20 of the 22 Mycobacterium species tested. The type strains of Mycobacterium tuberculosis and M. bovis were genetically highly related and could not be differentiated by this method. Of 194 biochemically identified human clinical strains, 178 (90%) were genetically identified within 3 h of the small-scale DNA extraction.
Kusunoki
S
S
Research and Development Center, Kobayashi Pharmaceutical Co., Osaka, Japan.
Ezaki
T
T
Tamesada
M
M
Hatanaka
Y
Y
Asano
K
K
Hashimoto
Y
Y
Yabuuchi
E
E
D010131:Aminosalicylic Acid / Q000378:metabolism; D001192:Arylsulfatases / Q000096:biosynthesis; D001426:Bacterial Proteins; D001560:Benzidines; D002863:Chromogenic Compounds; D003124:Colorimetry; D015342:DNA Probes; D009161:Mycobacterium / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D009525:Niacin / Q000096:biosynthesis; D009566:Nitrates / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D011136:Polysorbates / Q000378:metabolism; D019809:Streptavidin; D014510:Urease / Q000096:biosynthesis
D001426:Bacterial Proteins; D001560:Benzidines; D002863:Chromogenic Compounds; D015342:DNA Probes; D009566:Nitrates; D011136:Polysorbates; D009525:Niacin; C020515:N,N,N',N'-tetramethylbenzidine; D010131:Aminosalicylic Acid; D019809:Streptavidin; D001192:Arylsulfatases; D014510:Urease