Detection of leptospiral DNA by nucleic acid hybridisation with 32P- and biotin-labelled probes.
Dot hybridisation with 32P- and biotin-labelled probes prepared from leptospiral DNA was performed to develop a sensitive and specific diagnostic method for early infection with leptospires. The smallest amounts of leptospiral DNA that could be detected with 32P- and biotin-labelled probes was 1.5 pg and 5 pg, respectively, corresponding to about 750 and 2500 leptospires. Dot hybridisation with a 32P-labelled probe detected leptospiral DNA in sera from all of 14 experimentally infected golden hamsters. The smallest amount of leptospiral DNA detected in these experiments corresponded to about 2500 leptospires. In the test conditions described in this study, the sensitivity of dot hybridisation with a biotin-labelled probe was lower. Little cross-hybridisation was observed with unrelated DNAs which indicates that dot hybridisation could be a useful diagnostic method.
D000818:Animals; D001710:Biotin; D006224:Cricetinae; D004269:DNA, Bacterial / Q000032:analysis*; D007919:Leptospira / Q000235:genetics* / Q000302:isolation & purification; D008647:Mesocricetus; D009693:Nucleic Acid Hybridization; D010761:Phosphorus Radioisotopes
D004269:DNA, Bacterial; D010761:Phosphorus Radioisotopes; D001710:Biotin