Identification of Streptococcus species by randomly amplified polymorphic deoxyribonucleic acid fingerprinting.
Identification of Streptococcus species isolated from bovine milk by randomly amplified polymorphic DNA fingerprinting was evaluated. Bacterial DNA templates were prepared using a standardized method for randomly amplified polymorphic DNA amplification and previously evaluated arbitrary primers. Amplified DNA fragments were visualized by agarose gel electrophoresis and were analyzed by densitometry. Unidentified streptococci (n = 163) that were isolated from mammary secretions of dairy cows were evaluated. The DNA fingerprint patterns of unidentified bacteria were compared using a computerized database that contained DNA fingerprint patterns of test strains. Comparison with the API 20 Strep identification system (bioMérieux Vitek, Inc., Hazelwood, MO) and conventional biochemical tests showed that about 91% of isolates (148 of 163) were identified correctly by DNA fingerprinting. The sensitivity of the DNA fingerprinting technique was 90%, and the specificity was 92%. However, the DNA fingerprinting technique only identified 4 of the 11 species included in the study. Three of the 4 species, Streptococcus uberis, Streptococcus agalactiae, and Streptococcus dysgalactiae, represent the streptococci isolated most frequently from cows with mastitis. The other Streptococcus and Enterococcus species that were not identified by the DNA fingerprinting system are less frequently isolated as causative agents of mastitis. Expanding the DNA fingerprint database would likely increase the sensitivity and specificity of this technique.
Gillespie
B E
BE
Department of Animal Science, University of Tennessee, Knoxville 37901-1071, USA.
Jayarao
B M
BM
Oliver
S P
SP
D000818:Animals; D002417:Cattle / Q000382:microbiology*; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D016983:Enterococcus / Q000235:genetics; D005260:Female; D005784:Gene Amplification; D008414:Mastitis, Bovine / Q000382:microbiology; D008892:Milk / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013291:Streptococcus / Q000145:classification* / Q000235:genetics
D004269:DNA, Bacterial