Rapid and simplified protocol for isolation and characterization of leptospiral chromosomal DNA for taxonomy and diagnosis.

Journal of clinical microbiology ·Vol. 22 ·No. 4 ·1985-10-00 ·Pages 606-8

Le Febvre RB, Foley JW, Thiermann AB

Abstract

We have developed a rapid method for the isolation of leptospiral chromosomal DNA which yields DNA of a purity suitable for restriction endonuclease analysis. A small volume (15 to 20 ml) of an exponentially growing culture of leptospires yielded 2 to 4 micrograms of chromosomal DNA. In a 1-day protocol, the DNA was isolated, restricted with endonucleases, and fractionated on an agarose gel. Chromosomal DNA from dinger zones (visible subsurface zones of leptospiral growth) of first semisolid subcultures of field isolates was also isolated and characterized, thus greatly speeding up the diagnostic process.

MeSH Terms

Chromosomes, Bacterial DNA Restriction Enzymes DNA, Bacterial/analysis,isolation & purification Deoxyribonuclease EcoRI Leptospira interrogans/classification,genetics,growth & development Ultracentrifugation

Chemicals

DNA, Bacterial DNA Restriction Enzymes Deoxyribonuclease EcoRI

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Le Febvre R B

Foley J W

Thiermann A B

References (14)

14 references, click to expand

On the structure of the folded chromosome of Escherichia coli.
J Mol Biol. 1972 Nov 14;71(2):127-47 PMID: 4564477
Improved microtechnique for the leptospiral microscopic agglutination test.
Appl Microbiol. 1973 Jun;25(6):976-80 PMID: 4736794
RNA molecules bound to the folded bacterial genome stabilize DNA folds and segregate domains of supercoiling.
Cold Spring Harb Symp Quant Biol. 1974;38:31-41 PMID: 4598638
Studies on the folded chromosome of Escherichia coli.
Cold Spring Harb Symp Quant Biol. 1974;38:43-51 PMID: 4598640
Concentration of DNA solutions by extraction with 2-butanol.
Biochim Biophys Acta. 1975 Jan 6;378(1):18-21 PMID: 235311
Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5.
J Bacteriol. 1978 Jul;135(1):227-38 PMID: 97269
NUTRITION OF LEPTOSPIRA POMONA AND GROWTH OF 13 OTHER SEROTYPES: A SERUM-FREE MEDIUM EMPLOYING OLEIC ALBUMIN COMPLEX.
Am J Vet Res. 1965 Jan;26:39-44 PMID: 14266933
Identification of Leptospira serovars by restriction-endonuclease analysis.
J Med Microbiol. 1981 Feb;14(1):163-6 PMID: 6257907
The effect of spermidine on endonuclease inhibition by agarose contaminants.
Anal Biochem. 1981 Jul 15;115(1):42-5 PMID: 6272602
Differentiation of subtypes within Leptospira interrogans serovars Hardjo, Balcanica and Tarassovi, by bacterial restriction-endonuclease DNA analysis (BRENDA).
J Med Microbiol. 1982 Aug;15(3):331-8 PMID: 6288951
A rapid alkaline extraction method for the isolation of plasmid DNA.
Methods Enzymol. 1983;100:243-55 PMID: 6353143
Restriction endonuclease DNA analysis of Leptospira interrogans serovars icterohaemorrhagiae and hebdomadis.
J Clin Microbiol. 1984 Oct;20(4):808-10 PMID: 6092434
New method for classification of leptospiral isolates belonging to serogroup pomona by restriction endonuclease analysis: serovar kennewicki.
J Clin Microbiol. 1985 Apr;21(4):585-7 PMID: 2985648
Recovery of DNA from gels.
Methods Enzymol. 1980;65(1):371-80 PMID: 7374460

Article Info

Journal

Journal of clinical microbiology

Abbr.

J Clin Microbiol

ISSN

0095-1137

DOI

10.1128/jcm.22.4.606-608.1985

Published

1985-10-00

Pages

606-8

Language

English

Region

United States

NLM ID

7505564

PMCID

PMC268476

Subset

External Links

PubMed source DOI: 10.1128/jcm.22.4.606-608.1985 PMC Full Text View journal details

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Rapid and simplified protocol for isolation and characterization of leptospiral chromosomal DNA for taxonomy and diagnosis.

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