Rapid and specific detection of pathogenic Leptospira species by amplification of ribosomal sequences.
We have developed an assay for the detection of pathogenic Leptospira that is based on the polymerase chain reaction. With the combination of agarose gel electrophoresis and blotting, pathogenic Leptospira can be discriminated specifically from nonpathogenic Leptospira and other bacterial species. This method, based on the amplification of 16S ribosomal RNA sequences, is able to detect 10 leptospiral cells/mL in cattle urine samples and 100 leptospiral cells/mL in pig urine samples. Using this assay leptospires were detected in urine samples from cattle that were experimentally infected with Leptospira interrogans serovar hardjo type hardjobovis.
Wagenaar
J A
JA
Department of Bacteriology, School of Veterinary Medicine, Utrecht University, The Netherlands.
Segers
R P
RP
Van der Zeijst
B A
BA
D000818:Animals; D015373:Bacterial Typing Techniques; D001437:Bacteriuria / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology / Q000652:urine; D004269:DNA, Bacterial / Q000032:analysis*; D004275:DNA, Ribosomal / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D007921:Leptospira interrogans / Q000302:isolation & purification; D007922:Leptospirosis / Q000382:microbiology / Q000662:veterinary*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000382:microbiology / Q000652:urine; D014774:Virulence / Q000235:genetics
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S