Detection of bacteria by hybridization of rRNA with DNA-latex and immunodetection of hybrids.
A novel nucleic acid hybridization assay with a DNA probe immobilized on 1.25-micron-diameter latex particles was developed. Hybridization of the immobilized probe DNA with sample rRNA was complete in 10 to 15 min. Alkaline phosphatase-labeled anti-DNA-RNA was allowed to bind to the DNA-RNA hybrids on the latex particles. Then the latex was collected on a small glass fiber filter pad, and bound alkaline phosphatase was quantitated by reflectance rate measurement. The method detected a broad range of bacterial species and had a detection limit of 500 cells per assay. The assay was used to screen urine samples for bacteriuria and had a sensitivity of 96.2% compared with conventional culture at a decision level of greater than or equal to 10(4) CFU/ml. The hybridization method could have broad application to the detection of bacteria and viruses.
Miller
C A
CA
Miles, Inc., Elkhart, Indiana 46515.
Patterson
W L
WL
Johnson
P K
PK
Swartzell
C T
CT
Wogoman
F
F
Albarella
J P
JP
Carrico
R J
RJ
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001437:Bacteriuria / Q000175:diagnosis*; D055598:Chemical Phenomena; D002621:Chemistry; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008863:Microspheres; D009693:Nucleic Acid Hybridization; D011237:Predictive Value of Tests; D012329:RNA, Bacterial / Q000032:analysis*; D012335:RNA, Ribosomal / Q000032:analysis*
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012335:RNA, Ribosomal